KR20120101153A - Compositions and methods for preventing and treating mucositis and weight loss - Google Patents

Abstract

As a method for preventing and / or treating mucositis, (1) 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione or an analog, derivative, metabolite, prodrug or solvate thereof Or at least one compound selected from the group consisting of pharmaceutically acceptable salts, or (2) a cytokinin, in a therapeutically effective amount; And administering the composition to a subject in need thereof. In addition, pharmaceutical compositions comprising 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione and / or cytokinin compounds, and pharmaceuticals for treating and / or preventing mucositis Also provided is the use of the composition for the preparation. Between the non-talkie compound is N ⁶ - iso-pentenyl, and adenosine N ⁶ - may be selected from benzyl-adenosine, are not limited only to these. The composition may also further comprise a chemotherapeutic agent such as cisplatin. Methods and compositions are also provided for preventing and / or alleviating weight loss in a patient being treated for cancer.

Description

Compositions and methods for the prevention and treatment of mucositis and weight loss {COMPOSITIONS AND METHODS FOR PREVENTING AND TREATING MUCOSITIS AND WEIGHT LOSS}

The present invention provides methods and compositions for the prevention and / or treatment of mucositis. More specifically, the present invention provides oltipraz (5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione) or an analog or derivative thereof. Compositions comprising and / or compositions comprising N ⁶ -isopentenyl adenosine or an analog or derivative thereof, and their use for preventing and treating mucositis. The present invention also provides compositions and methods for preventing weight loss in a subject due to radiation therapy and for preventing and treating cachexia.

Mucositis means inflammation of the mucosa lining that covers the mucous membrane or digestive tract. This condition is caused by damage to the mucosa and can lead to the formation of ulcerative lesions. These lesions are extremely painful and can occur anywhere in the alimentary tract from the oral cavity to the anus, including the esophagus, stomach, small intestine, large intestine, and rectum.

Mucositis is a common side effect of chemotherapy or radiation therapy. The mucous membranes of the mouth and digestive tract are sensitive to both chemotherapy and radiation therapy. Chemotherapy agents used to treat the condition of cancer adversely affect normal cells, particularly those with high turnover rates, such as cells of oral epithelial tissue. Such radiation therapy causes cell death, which causes thinning, peeling, redness and inflammation of the mucosal lining, resulting in ulcers. Patients receiving chemotherapy show this symptom within four to five days after the start of treatment.

Mucositis associated with radiation therapy usually appears within 14 days of treatment and the symptoms last for 6 to 8 weeks.

Pathophysiology of mucositis is divided into five stages including initiation stage, message generation stage, signaling and amplification stage, ulceration stage and treatment stage. . Different steps are caused by different cytokines. The initiation step is initiated after chemotherapy or radiation therapy and produces free radicals that cause DNA damage. Subsequently, transcription factors such as NF-kB are produced to up regulate inflammatory cytokine production. This inflammation, mediated by cytokines such as IL-1 and TNF-alpha, leads to an ulceration stage.

The main clinical symptoms of mucositis are esophagitis (inflammation of the esophagus), dysphagia (difficulty to swallow), swallowing pain (odynophagia), and chest pain under the sternum (for mucositis induced by radiation). And chest pain behind the sternum (by combination therapy).

There is no effective treatment for mucositis. Current methods of treatment are to ameliorate symptoms and to maintain high levels of oral hygiene and include the use of topical analgesics such as lidocaine and mouthwashes such as chlorohexidine gluconate. Another therapeutic regimen involves the use of agents that reduce mucosal absorption of chemotherapy drugs, such as cryotherapy or allopurinol. Other therapeutic agents, such as glutamine or beta-carotene, reduce changes in epithelial cell proliferation. Still other therapies include laser treatment and antibiotics, as well as the use of cytokine-based therapeutic therapies such as Palipermin (trade name: Kepivance, Amgen), a human keratinocyte growth factor (KGF), and other inflammatory modulators.

None of the therapeutic approaches currently in use have proven to be fully effective in preventing and treating mucositis. Thus, in order to develop therapeutic therapies that can be used for the effective prevention and treatment of mucositis, substantial clinical research remains an open challenge. The treatment regimen is particularly useful in patients with cancer status who are planning to or are receiving treatment regimens for cancer such as chemotherapy and / or radiation therapy.

Cachexia refers to weight loss, muscle atrophy, fatigue, weakness, and severe food loss in a person who is not actively trying to lose weight. Cachexia is a symptom that manifests in various diseases, such as cancer, infectious diseases (such as tuberculosis, AIDS) and some autoimmune diseases, or addiction to drugs such as amphetamine or cocaine.

The inventors have surprisingly identified novel compounds and methods for preventing and treating mucositis in subjects undergoing radiation therapy. In particular, the inventors have found that many non-steroidal compounds can be used to prevent and treat mucositis unexpectedly. We also identified compositions that can be used to prevent and treat cachexia and to prevent weight loss in subjects undergoing cancer treatment of radiotherapy.

SUMMARY OF THE INVENTION

According to a first aspect of the present invention, as a method for preventing and / or treating mucositis, 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione (oltipraz) or an analog thereof Providing a therapeutically effective amount of a composition comprising one or more compounds selected from the group consisting of derivatives, metatabolites, prodrugs, solvates or pharmaceutically acceptable salts; And administering the composition to a subject.

According to a second aspect of the invention, 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione (oltipraz) or analogs, derivatives, metabolites, prodrugs, solvents thereof There is provided a composition comprising at least one compound selected from the group consisting of cargoes or pharmaceutically acceptable salts.

According to a third aspect of the invention, in the manufacture of a medicament for the prevention and / or treatment of mucositis, 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione (oltifra) J) or analogs, derivatives, metabolites, prodrugs, solvates or pharmaceutically acceptable salts thereof.

According to a fourth aspect of the invention there is provided a composition for the prevention and / or treatment of mucositis, the composition comprising 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione ( Oltipraz) or analogs, derivatives, metabolites, prodrugs, solvates or pharmaceutically acceptable salts thereof.

A fifth aspect of the invention relates to 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione (oltipraz) or analogs, derivatives, metabolites, prodrugs, solvates or It provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier with at least one compound selected from the group consisting of pharmaceutically acceptable salts.

In an embodiment of the aforementioned aspect of the invention, the metabolite of 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione is known as pyrrolopyrazine derivative metabolite 3 (also known as M3). )to be.

In an embodiment, the analog of 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione is a 1,2-dithiol-3-thione-based compound, for example ane Tol trithion ((5- (p-methoxyphenyl) -3H-1,2-dithiol-3-thio) (also known as anetitol trithiona or SONICUR ^™ ).

In an embodiment, the 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione compound is 1,2 dithiol-3-thione (D3T) or an analog thereof. Typically 1,2 dithiol-3-thione analogs have the formula:

Wherein, in the case of a 5-substituted analog,

R ₁ is H, R ₂ is phenyl and X is S; or

R ₁ is H, R ₂ is 4-methoxyyl, X is S; or

R ₁ is H, R ₂ is 2-pyrazinyl, and X is 0; or

R ₁ is H, R ₂ is 2- (5,6-dimethyl) pyrazinyl and X is S; or

For 5-substituted-4-methyl analogs,

R ₁ is CH ₃ , R ₂ is 2-pyridyl and X is S; or

R ₁ is CH ₃ , R ₂ is 3-pyridyl and X is S; or

R ₁ is CH ₃ , R ₂ is 4-pyridyl and X is S; or

R ₁ is CH ₃ , R ₂ is 3-pyridazinyl and X is S; or

R ₁ is CH ₃ , R ₂ is 2-thiofuranyl and X is S; or

R ₁ is CH ₃ , R ₂ is 2- (2-pyrazinyl) ethylene and X is S.

For 4-substituted-5- (2-pyrazinyl) analogs,

R ₁ is CH ₃ , R ₂ is 2-pyrazinyl and X is S; or

R ₁ is CH ₃ , R ₂ is 2-pyrazinyl and X is 0; or

R ₁ is CH ₂ 0H, R ₂ is 2-pyrazinyl and X is S; or

R ₁ is CH ₂ CH ₃ , R ₂ is 2-pyrazinyl and X is S; or

R ₁ is (CH ₂ ) ₃ CH ₃ , R ₂ is 2-pyrazinyl and X is S.

For other analogues,

R ₁ is CO ₂ C ₂ H ₅ , R ₂ is 2-pyridyl and X is S; or

R ₁ is CO ₂ C ₂ H ₅ , R ₂ is 4-pyridyl and X is S; or

R ₁ is CI, R ₂ is [4- (2-propyl) phenyl] amino and X is S; or

R ₁ is CI, R ₂ is [4- (2-propyl) phenyl] amino and X is 0; or

R ₁ is CH ₂ CO ₂ C ₂ H ₅ , R ₂ is 5-pyrimidyl and X is S; or

R ₁ is CH ₂ CON [CH (CH ₃ ) ₂ H] ₂ , R ₂ is 5-pyrimidyl and X is S; or

R ₁ is phenethyl, R ₂ is 3-pyridazinyl and X is S; or

R ₁ is H, R 2 is 4-pyridyl, and X is NO— (CH ₂ ) ₃ N (CH ₃ ) ₂ ; or

R ₁ is (CH ₂ ) ₃ CH ₃ , R ₂ is 3- (6-dimethylamino) pyridazinyl) and X is S.

In another embodiment, R ₁ is fluorine or bromine and R ₂ and X are selected from the substituents listed above.

In an embodiment, the compounds of the present invention are formulated with or co-administered with carboxymethyl cellulose (CMC).

The inventors surprisingly 5- [2-pyrazinyl] -4-methyl- when 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione is co-administered with carboxymethyl cellulose. It was found that the toxicity associated with 1,2-dithiol-3-thione administration was significantly reduced. In particular, the inventors have found that, when formulated with CMC, 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione in an amount of up to 2000 mg / kg without causing serious toxicity It was confirmed that can be administered to the subject. Without wishing to be bound by theory, when formulated with carboxymethyl cellulose, 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione is not absorbed into the bloodstream and the digestive tract It is thought to form an effective lining of the digestive tract, which combines with the outer wall of the gut, protecting it from damage such as gastrointestinal damage.

In an embodiment, the 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione compound is an amino acid containing sulfur, for example cysteine or an analog, derivative, salt or solvate thereof And co-administered or formulated together.

5- [2-pyrazinyl] -4-methyl-1,2-dithiine when orally administered cysteine with 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione There was a significant increase in both bioavailability and bioavailability of all-3-thione (Hassan M. Ali et aI., 1984; Chemotherapy 30: 255-261).

In an embodiment, the 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione compound consists of cisplatin, dexamethasone and 5-fluorouracil, It may be co-administered or formulated with a chemotherapeutic agent, which may be selected from the group which is not limited thereto.

We have also surprisingly found that cytokinin compounds can be used for the prevention and / or treatment and / or amelioration of mucositis. Accordingly, the present invention further encompasses the use of methods, compositions and cytokinin compounds for the treatment, amelioration and / or prevention of mucositis.

Thus, in another aspect of the present invention, a method of preventing and / or treating mucositis is provided, wherein the method comprises one or more cytokinin compounds or pharmaceutically acceptable salts or solvates thereof. Providing a composition of the same, and administering the composition to a subject in need thereof.

In another aspect, the present invention provides a composition comprising at least one cytokinin compound or a pharmaceutically acceptable salt or solvate thereof.

In another aspect, the invention provides the use of a composition comprising at least one cytokinin compound or a pharmaceutically acceptable salt or solvate thereof in the manufacture of a medicament for the treatment and / or prophylaxis of mucositis.

In another aspect, the invention provides a composition comprising one or more cytokinin compounds or pharmaceutically acceptable salts or solvent compounds thereof for use in the treatment and / or prevention of mucositis.

In another aspect, the present invention provides a pharmaceutical composition comprising at least one cytokinin compound or a pharmaceutically acceptable salt or solvent compound thereof together with a pharmaceutically acceptable carrier or diluent.

In an embodiment, the cytokinin compound is N ⁶ -isopentenyl adenosine (IPA) or an analog, derivative, metabolite, prodrug, solvent compound or salt thereof.

In another embodiment, the cytokinin compound is N ⁶ -benzyl adenosine (IPA) or an analog, derivative, metabolite, prodrug, solvent compound or salt thereof.

In yet another embodiment, the cytokinin compound is selected from the group including but not limited to kinetin, zeatin and benzyl adenine. In particular, the cytokinin compounds are 6- (substituted amino) purines, for example kinetin (6- (furfuryl) aminopurine), zeatin (6- (3-hydroxymethyl, 3-methylallyl), amino Purine, 6- (3,3-dimethylallyl) aminopurine, 6- (benzyl) aminopurine, 6- (phenyl) aminopurine, 6- (n-alkyl) aminopurine, wherein the alkyl group is 4, 5 or 6 Carbon atoms) and 6- (cyclohexyl) methylaminopurine In one embodiment, the 6- (substituted amino) purine cytokinin is from about 0.01% (w / v) to about 0.5% (w / v), preferably with a physiologically acceptable carrier or diluent at a concentration of about 0.1% (w / v).

In an embodiment, the cytokinin compound may be co-administered or formulated with chemotherapeutic agents such as cisplatin, dexamethasone and 5-fluorouracil.

In an embodiment, the cytokinin compound is co-administered or formulated with carboxymethyl cellulose (CMC).

The present inventors have surprisingly found that a typical cytokinin compound, N ⁶ -isopentenyl adenosine, can be significantly reduced in toxicity by administration with carboxymethyl cellulose. Specifically, the inventors have been shown that administering 50 to 100 mg / kg of the cytokinin compound without CMC causes toxicity in the liver, whereas when formulated with CMC, the subject can administer the cytokinin compound to 2000 mg / kg. Also found safe. Without wishing to be bound by theory, it is believed that when administered with carboxymethyl cellulose, cytokinin compounds do not absorb into the bloodstream but form a membrane in the digestive tract, which serves to protect against damage such as gastrointestinal damage.

In another aspect, the invention provides one or more cytokinin compounds or pharmaceutically acceptable salts or solvates thereof and 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione Or a combined medicament comprising an analog, derivative, metabolite, prodrug, solvate or pharmaceutically acceptable salt thereof.

The present invention also provides a pharmaceutical composition for use in the prevention or treatment of mucositis, comprising the combination medicament together with one or more pharmaceutically acceptable carriers or diluents.

The present invention also provides the use of the above-mentioned combination medicament or a pharmaceutical composition comprising the same in the practice of the method for preventing and / or treating mucositis.

According to another aspect of the present invention, there is provided a method for preventing and / or treating mucositis, wherein the method includes 3H-1,2-dithiol-3-thione, anetitol trithione (5- (4-meth). Methoxyphenyl) -3H-1,2-dithiol-3-thione), ADT, ADO, 1,2-dithiol-3-thione, 1,2-dithiolane, 1,3-dithiol-2-thione , Malolate, 4- (3,5-diisopropyl-4-hydroxyphenyl) -1,2-dithiol-3-thione; 4- (3,5-di-t-butyl-4-hydroxyphenyl) -1,2-dithiol-3-thione; 4- [3,5-bis (1,1-dimethylpropyl) -4-hydroxyphenyl] -1,2-dithiol-3-thione; 4- [3,5-bis (1,1-dimethylbutyl) -4-hydroxyphenyl] -1,2-dithiol-3-thione; 4- [3,5-bis (1,1,3,3-tetramethylbutyl) -4-hydroxyphenyl] -1,2-dithiol-3-thione; 4- [3,5-bis (1-methylcyclohexyl) -4-hydroxyphenyl] -1,2-dithiol-3-thione; 4- [3,5-bis (1,1-dimethylbenzyl) -4-hydroxyphenyl] -1,2-dithiol-3-thione; 4- (3t-butyl-4-hydroxy-S-isopropylphenyl) -1,2-dithiol-3-thione; 4- (3t-butyl-4-hydroxy-5-methylphenyl) -1,2-dithiol-3-thione; 4- [3 (1,1-dimethylpropyl) -4-hydroxy-5-isopropylphenyl] -1,2-dithiol-3-thione; 4- [3 (1,1-dimethylbenzyl) -4-hydroxy-5-isopropylphenyl] -1,2-dithiol-3-thione; 5-benzylthio-4- (3,5-di-t-butyl-4-hydroxyphenyl) -1,2-dithiol-3-thione; 5-benzylthio-4- [3,5-bis (1,1-dimethylpropyl) -4-hydroxy-phenyl] -1,2-dithiol-3-thione; 5-hexylthio-4- (3,5-di-t-butyl-4-hydroxyphenyl) -1,2-dithiol-3-thione; 5-hexylthio-4- [3,5-bis (1,1-dimethylbutyl) -4-hydroxy-phenyl] -1,2-di-thiol-3-thione; 5-octadecylthio-4- (3,5-di-t-butyl-4-hydroxyphenyl) -1,2-dithiol-3-thione; 5-octadecylthio-4- [3,5-bis (1,1-dimethylbenzyl) -4-hydroxyphenyl] -1,2-dithiol-3-thione; 5-allylthio-4- (3,5-di-t-butyl-4-hydroxyphenyl) -1,2-dithiol-3-thione; 5-cyclohexylthio-4- (3,5-di-t-butyl-4-hydroxyphenyl) -1,2-dithiol-3-thione; And at least one compound selected from the group consisting of 4- (3,5-di-sec-butyl-4-hydroxyphenyl) -1,2-di-thiol-3-thione, in a therapeutically effective amount Providing and administering said composition to a subject in need thereof.

Another aspect of the invention, 3H-1,2-dithiol-3-thione, anetol trithione (5- (4-methoxyphenyl) -3H-1,2-dithiol-3-thione), ADT, ADO, 1,2-dithiol-3-thione, 1,2-dithiolane, 1,3-dithiol-2-thione, malolate, 4- (3,5-diisopropyl-4- Hydroxyphenyl) -1,2-dithiol-3-thione; 4- (3,5-di-t-butyl-4-hydroxyphenyl) -1,2-dithiol-3-thione; 4- [3,5-bis (1,1-dimethylpropyl) -4-hydroxyphenyl] -1,2-dithiol-3-thione; 4- [3,5-bis (1,1-dimethylbutyl) -4-hydroxyphenyl] -1,2-dithiol-3-thione; 4- [3,5-bis (1,1,3,3-tetramethylbutyl) -4-hydroxyphenyl] -1,2-dithiol-3-thione; 4- [3,5-bis (1-methylcyclohexyl) -4-hydroxyphenyl] -1,2-dithiol-3-thione; 4- [3,5-bis (1,1-dimethylbenzyl) -4-hydroxyphenyl] -1,2-dithiol-3-thione; 4- (3t-butyl-4-hydroxy-S-isopropylphenyl) -1,2-dithiol-3-thione; 4- (3t-butyl-4-hydroxy-5-methylphenyl) -1,2-dithiol-3-thione; 4- [3 (1,1-dimethylpropyl) -4-hydroxy-5-isopropylphenyl] -1,2-dithiol-3-thione; 4- (3 (1,1-dimethylbenzyl) -4-hydroxy-5-isopropylphenyl] -1,2-dithiol-3-thione; 5-benzylthio-4- (3,5-di- t-butyl-4-hydroxyphenyl) -1,2-dithiol-3-thione; 5-benzylthio-4- [3,5-bis (1,1-dimethylpropyl) -4-hydroxy-phenyl ] -1,2-dithiol-3-thione, 5-hexylthio-4- (3,5-di-t-butyl-4-hydroxyphenyl) -1,2-dithiol-3--thione; 5-hexylthio-4- [3,5-bis (1,1-dimethylbutyl) -4-hydroxy-phenyl] -1,2-dithiol-3-thione; 5-octadecylthio-4- ( 3,5-di-t-butyl-4-hydroxyphenyl) -1,2-dithiol-3-thione; 5-octadecylthio-4- [3,5-bis (1,1-dimethylbenzyl) 4-hydroxyphenyl] -1,2-dithiol-3-thione; 5-allylthio-4- (3,5-di-t-butyl-4-hydroxyphenyl) -1,2-dithiol 3-thione; 5-cyclohexylthio-4- (3,5-di-t-butyl-4-hydroxyphenyl) -1,2-dithiol-3-thione; and 4- (3,5- With one or more compounds selected from the group consisting of di-sec-butyl-4-hydroxyphenyl) -1,2-dithiol-3-thione It provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier or diluent.

In another aspect, the invention provides a method of preventing and / or preventing weight loss in a subject being treated for cancer, the method comprising: 5- [2-pyrazinyl] -4-methyl-1,2 -Providing a therapeutically effective amount of a composition comprising at least one compound selected from the group consisting of dithiol-3-thione or analogs, derivatives, metabolites, prodrugs, solvates or pharmaceutically acceptable salts thereof And administering said composition to a subject.

In another aspect, the invention provides 5- [2-pyrazinyl] -4-methyl-1,2-dithiol- in the manufacture of a medicament for preventing and / or preventing weight loss in a subject being treated for cancer. Provided is a use of a composition comprising one or more compounds selected from the group consisting of 3-thione or analogs, derivatives, metabolites, prodrugs, solvates or pharmaceutically acceptable salts thereof.

In another aspect, the invention provides 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione to prevent and / or prevent weight loss in a subject being treated for cancer. Or analogs, derivatives, metabolites, prodrugs, solvates or pharmaceutically acceptable salts thereof.

In an embodiment, the cancer treatment is chemotherapy, radiation therapy or a combination therapy thereof.

In another aspect of the invention, the invention provides a method for preventing and / or treating cachexia, the method comprising: 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3- Providing a therapeutically effective amount of a composition comprising one or more compounds selected from the group consisting of thio or analogs, derivatives, metabolites, prodrugs, solvates or pharmaceutically acceptable salts thereof and to a subject in need thereof Administering the composition.

In another aspect of the invention, the invention provides 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione or an analog thereof in the manufacture of a medicament for cachexia prevention and / or treatment. Provided is a use of a composition comprising at least one compound selected from the group consisting of derivatives, metabolites, prodrugs, solvates or pharmaceutically acceptable salts.

In another aspect of the invention, the invention provides 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione or an analog thereof for use in cachexia prevention and / or treatment, Provided are compositions comprising derivatives, metabolites, prodrugs, solvates or pharmaceutically acceptable salts.

In an embodiment of the aforementioned aspect of the invention, the metabolite of 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione is known as pyrrolopyrazine derivative metabolite 3 (also known as M3). )to be.

In an embodiment, 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione is selected from anetitol trithion (anetitol trithion or SONICUR ^™). Also known as).

In an embodiment, the 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione compound is co-administered or formulated with cysteine or an analog, derivative salt or solvate thereof.

In an embodiment, the 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione compound is selected from the group consisting of but not limited to cisplatin, dexamethasone and 5-fluorouracil Or in combination with a chemotherapeutic which may be used.

In an embodiment, the subject has been treated for cancer by chemotherapy, radiation therapy or a combination thereof.

In an embodiment, the cachexia is cancer cachexia.

Without wishing to be bound by theory, the present invention is based, in part, on the inventors' unexpected finding that thinning and ulceration of the mucous membranes of the subject's digestive tract (GI) are prevented when the subject is treated with the composition of the present invention.

The term "mucositis" as used herein is intended to include alimentary mucositis. In an embodiment, gastrointestinal mucositis includes oral mucositis and / or enteritis (intestinal inflammation, in particular inflammation of the small intestine).

In an embodiment, gut mucositis includes esophagitis (inflammation of the esophagus), pharyngeal mucositis, stomatitis (inflammation of the stomach) and / or proctitis (inflammation of the rectum).

In an embodiment, the methods and uses of the present invention comprise administering at least one compound of the present invention to at least one digestive tract region in a therapeutically effective amount for a subject having or at risk of developing mucositis. In an embodiment, one or more compounds can be administered to one or more digestive tract areas.

As used herein, the term "cachexia" refers to unwanted weight loss in a person who does not actively strive for weight loss. As used herein, the expression "prevention and / or prevention of weight loss" and similar expressions includes no change in body weight and / or no increase in body weight.

In an embodiment, the compositions, methods and uses extend to the prevention of mucositis and / or weight loss in subjects undergoing radiation therapy and / or chemotherapy. In an embodiment, one or more compounds of the present invention are administered to a subject prior to conditioning for myeloablative radiotherapy and / or chemotherapy in preparation for transplantation of autologous or heterologous hematopoietic stem cells May be administered.

In an embodiment, the present invention provides compositions and methods for mucositis prophylaxis and / or treatment and / or weight loss in subjects who have received or are going to receive mucosal toxic chemotherapy with mucositis-inducing agents.

In an embodiment, the present invention provides a method for preventing and / or treating mucositis in a subject who is receiving or will receive radiation therapy, with or without adjuvant chemotherapy, and has cancer in the head and / or neck. To provide a composition.

In an embodiment, mucositis and / or weight loss occurs in a subject exposed to chemical injury, biological injury, radiation, or a combination thereof. Radiation exposure may result from radiation therapy, for example chemotherapy, radiation therapy, or the like, or may be caused by accidental radiation exposure or radiation exposure by terrorist attack. The compositions, methods and uses of the present invention have additional uses in connection with administration to a subject before or after space travel, to prevent, treat or ameliorate mucositis and / or weight loss.

In an embodiment, the method or use of the invention can be carried out before the subject suffers an injury that can cause or cause the progression of mucositis and / or weight loss.

In another embodiment, the methods or uses of the present invention may be carried out after the subject is injured, but before the onset and development of mucositis and / or weight loss in the subject.

In another aspect, the methods or uses of the invention may be performed on a subject after mucositis in the subject and / or weight loss of the subject occurs.

The compositions and methods of the present invention can be used in combination with other treatment regimens to prevent and / or treat mucositis and / or weight loss. For example, a composition comprising 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione and / or N ⁶ -isopentenyl adenosine and any one or more pharmaceutically acceptable The carrier may be administered in combination with one or more additional therapeutic agents that have a prophylactic and / or therapeutic effect on the occurrence or progression of mucositis or ameliorate one or more symptoms with respect to reducing mucositis or weight loss. . Non-limiting examples of such additional therapeutic agents include laser therapy, cryotherapy, antibiotics, cytokine-based therapeutic therapies such as palipermine (trade name: Kepivance, Amgen), a human keratinocyte growth factor (KGF), and IL- 11, other cytokine inflammation modulators such as TGF and GM-CSF.

In addition, the compositions of the present invention may be used in the manufacture of a combination medicament comprising one or more compounds of the present invention in combination with a chemotherapeutic agent.

Chemotherapeutic agents suitable for use with the compositions of the present invention include, for example, mitosis inhibitors such as vinblastine; Alkylating agents such as cisplatin, carboplatin and cyclophosphamide; Inhibitors of microtubule assembly, such as paclitaxel or other taxanes; Metabolic antagonists such as 5-fluorouracil, capecitabine, cytosine arabinoside and hydroxyurea; Intercalating antibiotics such as adriamycin and bleomycin; Immunostimulants such as trastuzumab; DNA synthesis inhibitors such as gemcitabine; Enzymes such as asparaginase; Topoisomerase inhibitors such as etoposide; Biological response modifiers such as interferon; And anti-hormonal agents such as, for example, antioestrogens such as tamoxifen, or (4'-cyano-3- (4-fluorophenylsulfonyl) -2-hydroxy-2-methyl-3 Antiandrogens such as'-(trifluoromethyl) -propionanilide and other therapeutic agents and for example DeVita, VT, Jr., Hellmann, S., Rosenberg, SA; in: Cancer: Principles & Practice of Oncology, 5th ed., Lippincott-Raven Publishers (1997).

The present invention also provides a method for administering the combination medicine. In an embodiment, compounds of the invention and chemotherapeutic agents are provided, respectively, continuously, simultaneously or through different routes of administration. In addition, the compounds and chemotherapeutic agents may be in the same or different forms, for example solid and liquid forms. The method may comprise administering a compound of the present invention simultaneously with a chemotherapeutic agent. In an embodiment, a compound of the present invention can be administered to a subject continuously with a chemotherapeutic agent. When administered simultaneously, in one embodiment, the compound of the present invention may be administered before the chemotherapeutic agent. In another embodiment, the compounds of the present invention may be administered later than the chemotherapeutic agent. In one embodiment, the chemotherapeutic agent is provided separately from the compound of the present invention.

In an embodiment, the chemotherapeutic agent and the compound of the present invention are administered in combination. Co-administration means that these components are administered together as a composition or as part of the same single dose. As used herein, the term “combination administration” also means that the components are administered separately but as part of a treatment regimen or treatment program. In one embodiment, the components are administered to the subject at the same time. However, the components may also be administered separately as discrete dosage or dosage forms. When the components are administered separately, the combined administration of the components does not limit the time, frequency, dosage, or order of administration of the components.

5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione (oltipraz, 4-methyl-5 (2-pyrazinyl) -3H-1,2-dithiol-3 The structure of -thione or 5- (2-pyrazinyl) -4-methyl-1,2-dithiol-3-thione) is represented by the following general formula (1).

[Formula 1]

In an embodiment, 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione forms a chelate or complexes with one or more divalent or trivalent radioactive metal ions By forming a, the divalent or trivalent radioactive ions are redistributed or sequestering in the subject cell or tissue, thereby limiting the capacity of the ions to participate in unwanted tissue destruction. Divalent or trivalent metal ions include iron, copper, nickel, calcium, magnesium, manganese, cadmium, lead, aluminum, silver, cobalt, iodine, zinc, mercury, cesium, uranium, selenium, protaxinium, thorium, rhodium and But selected from the group consisting of cerium ions or radicals.

Without wishing to be bound by theory, the inventors have indicated that the therapeutic and / or prophylactic treatment of 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione in connection with the prevention or treatment of mucositis. The effect was attributed to increased expression of glutathione (GSH), glutathione reductase and / or glutathione-S-transferase.

Cytokinins are known plant growth hormones that have activity in cell division, cell growth and differentiation and other physiological processes. Cytokine is involved in the growth of explants of plant tissues and the enhancement of cell division when cultured in standard media, containing vitamins, mineral salts and sugars as well as auxin (another kind of plant hormone). In particular, cytokinin is active in the process of regulating disease resistance, stress resistance, drought resistance, lodging resistance, aging delay, apical dominance and assimilation partitioning in plants. (Werner et al., Proc. Natl. Acad. Sci, 98 (18) 10487 10492 (2001), Haberer et al., Plant Physiol., 128, pp. 354 362 (2002).

The term "cytokinin" as defined herein means a compound that is a plant growth substance (plant hormone) that is involved in cell growth and differentiation as well as other processes. In particular, the term is also referred to as "adenine cytokinin" and includes cytokinins, including kinetin, zeatin and benzyl adenine. The term further includes "phenylurea cytokinin", having a different chemical composition, such as N, N'-diphenylurea, but having a biological activity similar to adenine cytokinin.

Suitable cytokinin compounds for use in the foregoing aspects of the invention are defined as Formula 2 below.

[Formula 2]

Where

R ₁ = H, R ₂ = CH ₃ , R ₃ = CH ₃ And R ₄ = H, or

R ₁ = H or CH ₃ S, and R ₄ is as follows:

및

And

R ₅ = CH ₃ , CI, OH or monophosphate group,

R ₆ = CH ₃ , CH ₂ OH or Cl,

R ₇ = H or Br, or

R ₁ = H and R ₄ is as follows:

X ₁ And X ₂ is independently selected from H, methyl group, ethyl group, hydroxyl group, halogen and carboxyl group; or

R ₄ is one of the following formulas:

,또는또는

,or or

Where R ₈ is

또는 (CH₂)₇CH_{3 ,} 이고,

Or (CH _{2) 7} CH _3, and,

R ₂ = OH and R ₃ = OH, monophosphate group, diphosphate group or triphosphate group, or

R ₂ and R ₃ are joined to each other to form a 3 ′, 5′-cyclic monophosphate derivative or a physiologically acceptable salt of any such compound.

Formula 2 is identified as "poly N ⁶ -isopentenyl adenosine" and is preferably used to mean all such compounds and salts, as well as polymers of IPA comprising 2 to 3 monomers.

The formulas listed below represent the chemical groups of R ₁ to R ₄ of the preferred compounds la to lu of formula (2).

[Formula la]:

In Formula 2, R ₁ = H, R ₂ = OH, R ₃ = OH, and R ₄ is as follows:

The compound is known as N ^6- (Δ ² -isopentenyl) adenosine.

Formula lb:

In Formula 2, R ₁ = H, R ₂ = OH, R ₃ = monophosphate, and R ₄ is as follows:

The compound is known as N ^6- (Δ ² -isopentenyl) adenosine-5'-monophosphate.

Formula lc:

In Formula 2, R ₁ = H, R ₂ and R ₃ combine to form a 3 ', 5'-cyclic monophosphate derivative, wherein R ₄ is as follows:

The compound is known as N ^6- (Δ ² -isopentenyl) adenosine-5'-cyclic monophosphate.

[Formula ld]:

In Formula 2, R ₁ = H, R ₂ = OH, R ₃ = OH, and R ₄ = CH ₂ C ₆ H ₆ .

This compound is known as N ⁶ -benzyladenosine.

[Formula le]:

In Formula 2, R ₁ = H, R ₂ = OH, R ₃ = monophosphate, and R ₄ = CH ₂ C ₆ H ₆ .

This compound is known as N ⁶ -benzyladenosine-5'-monophosphate.

Formula lf:

In Formula 2, R ₁ = H, R ₂ and R ₃ combine to form a 3 ', 5'-cyclic monophosphate derivative, and R ₄ = CH ₂ C ₆ H ₆ .

This compound is known as N ⁶ -benzyladenosine-3 ', 5'-cyclic monophosphate.

[Formula lg]:

In Formula 2, R ₁ = H, R ₂ = OH, R ₃ = OH, and R ₄ is as follows:

This compound is known as furfuryl adenosine.

[Formula lh]:

In Formula 2, R ₁ = H, R ₂ = OH, R ₃ = monophosphate, and R ₄ is as follows:

This compound is known as N ⁶ -fupuryladenosine-5'monophosphate.

[Formula li]:

In Formula 2, R ₁ = H, R _2, and R ₃ combine to form a 3 ′, 5′-cyclic monophosphate derivative, and R ₄ is as follows:

The compound is known as N ⁶ -furfuryl adenosine-3 ', 5'-cyclic monophosphate.

[Formula lj]:

In Formula 2, R ₁ = H, R ₂ = OH, R ₃ = OH, and R ₄ is as follows:

This compound is known as N- (purin-6-ylcarbamoyl) -O-chloroaniline ribonucleoside.

[Formula lk]:

In Formula 2, R ₁ = H, R ₂ = OH, R ₃ = monophosphate, and R ₄ is as follows:

This compound is known as N- (purin-6-ylcarbamoyl) -O-chloroaniline ribonucleoside-5'-monophosphate.

[Formula ll]:

In Formula 2, R ₁ = H, R ₂ = OH, R ₃ = OH, and R ₄ is as follows:

The compound is known as N ⁶ -adamantyladenosine.

Formula lm:

In Formula 2, R ₁ = H, R ₂ = OH, R ₃ = monophosphate, and R ₄ is as follows:

The compound is known as N ⁶ -adamantyladenosine-5'-monophosphate.

Formula ln:

In Formula 2, R ₁ = H, R ₂ = OH, R ₃ = OH, and R ₄ is as follows:

This compound is known as N- (purin-6-ylcarbamoyl) -n-octylamine ribocucleoside.

[Formula lo]

In Formula 2, R ₁ = H, R ₂ = OH, R ₃ = monophosphate, and R ₄ is as follows:

The compound is known as N- (purin-6-ylcarbamoyl) -n-octylamine ribocucleoside-5'-monophosphate.

[Formula lp]

In Formula 2, R ₁ = H, R _2, and R ₃ combine to form a 3 ′, 5′-cyclic monophosphate derivative, and R ₄ is as follows:

This compound is known as N- (purin-6-ylcarbamoyl) -n-octylamine ribonucleoside-3 ', 5'-monophosphate.

Formula lq]

In Formula 2, R ₁ = CH ₃ S, R ₂ = OH, R ₃ = OH, and R ₄ is as follows:

The compound is known as N ^6- (Δ ² -isopentyl) -2-methylthioadenosine.

[Formula lr]

In Formula 2, R ₁ = H, R ₂ = OH, R ₃ = OH, and R ₄ is as follows:

The compound is known as N ^6- (4-hydroxy-3-methyl- trans -2-butenyl) -adenosine.

[Formula ls]

In Formula 2, R ₁ = H, R ₂ = OH, R ₃ = OH, and R ₄ is as follows:

The compound is known as N ^6- (3-chloro- trans -2-butenyl) adenosine.

[Formula lt]

In Formula 2, R ₁ = H, R ₂ = OH, R ₃ = OH, and R ₄ is as follows:

The compound is known as N ^6- (3-chloro- cis -2-butenyl) adenosine.

[Formula lu]

In Formula 2, R ₁ = H, R ₂ = CH ₃ , R ₃ = CH ₃ , and R ₄ = H.

The present invention extends to one or more metabolites of the compound of formula (2). For example, preferred metabolites include N ^6- (Δ ² -isopentenyl) adenine, 6-N- (3-methyl-3-hydroxybutylamino) purine, adenine, hypoxanthine, uric acid ) And methylated xanthine.

Without wishing to be bound by theory, it is believed that the cytokinin compounds of formula (2) enhance the production of these enzymes by these cells after depletion of phase II detoxification enzymes by radiation exposure. . Phase II detoxification enzymes include glutathione-S transferase, gamma-glutamylcysteine synthetase, glutathione deductase, glutathione peroxidase, epoxide hydrase, AFB-1 aldehyde reductase, glucuronyl reductase, Glucose-6-phosphate dehydrogenase, UDP-glucuronyl transferase and AND (P) H: quinone oxidoreductase.

In one aspect, the invention extends to compositions, methods and uses comprising one or more of the following compounds and pharmaceutically acceptable salts:

ADT with the following structural formula:

ADO with the following structural formula:

1,2-dithiol 3-thione having the structure

Lipoamide (1,2-dithiolane) having the structure

1,3-dithiol 2-thione having the structure

[1,2] dithiol [4,3-c] -1,2-dithiol-3,6-dithione having the structure

In one aspect, the invention extends to compositions, methods and uses comprising one or more of the following compounds:

1,2-dithiolane class 1 compounds having the general formula:

1,2-dithiolane class 2 compounds having the general formula:

1,2-dithiolane class 3 compounds having the general formula:

1,2-dithiolane class 4 compound having the general formula:

Wherein Z = S, O, NR, R ₂ , CR ₂ , and Z is selected arbitrarily and independently for all the above classes. In this case, R includes H, alkyl (C1-C5), alkoxy (C1-C5), alkoxycarbonyl (C1-C5). R ₂ may form a spiro ring about the carbon atom of the ring.

For thiolane classes, the carbon atoms of the ring may be double substituted.

R ₁ -R ₄ are the major ring substituents for all classes, and optionally and independently H, alkyl, aryl, heterocyclic, halogen, alkoxycarbonyl (C1-C5) or carboxyl to include a wide variety of substituents Must include.

R ₁ , R ₂ , R ₃ , and R ₄ may form a spiro ring around the carbon atom to which they are attached or may form a ring bonded or bridged to an adjacent carbon atom.

The following definitions are for most of the compounds described herein.

Alkyl is C1-C10 saturated or unsaturated straight or branched chain herein and may be optionally substituted with one or multiple substituents by the following substituents: halogen, alkyl (C1-C5), hydroxyl, alkoxy (C1-C5) , Alkoxycarbonyl (C1-C5), carboxyl, amido, alkylamido (C1-C5), amino, mono and dialkyl amino (C1-C5), alkyl carbamoyl (C1-C5), thiols, Alkylthio (C1-C5) or benzenoid aryl.

Aryl is defined herein as any optionally single or multiple substituted benzenoid groups (C6-C14). The substituents are defined below.

Heterocyclic radicals are any 4, 5, or 6-membered optionally substituted heterocyclic rings, which may be substituted or unsubstituted and the ring may contain 1-3 N, O or S atoms and the remainder It contains carbon atoms.

Substituents for aryl or heterocyclic radicals include halogen, alkyl (C1-C5), hydroxyl, alkoxy (C1-C5), alkoxycarbonyl (C1-C5), carboxyl, amido, alkylamido (C1 -C5), amino, mono and dialkyl amino (C1-C5), alkyl carbamoyl (C1-C5), thiols, alkylthio (C1-C5) or benzenoid aryl cyano, nitro, halo alkyls, alkyl Sulfonyl (C1-C5), sulfonates. Two of these substituents may be part of a bonded ring, which ring may be a substituted or unsubstituted heterocyclic or carbocyclic ring.

Where

X is

(R5 is a C1-C6 alkyl or aryl group)

Z and Z 'are electron-attracting groups such as ester or cyano groups.

A is a> C = N-OH group or> C = N-OR ₃ Group (R ₃ is selected from hydroxyl, amino, chloro and C1-C4 alkoxy groups, aryl (C1-C6 alkyl) groups, (C1-C6 alkyl) carbonyl groups and aryl (C1-C6 alkyl) carbonyl groups).

A may be selected from> C═O groups,> C═NR ₄ (R ₄ is a C1-C6 alkyl group or an aryl group), and a CHOH group.

R ₁ and R ₂ are hydrogen, halogen, nitro group, nitro group, thiocano group, C1-C6 alkyl group, C2-C6 alkenyl group, aryl group, aryl (C1-C6 alkyl) group, aryl (C2-C6 alkenyl ), Carboxyl, (C1-C6 alkyl) carbonyl group, arylcarbonyl group, (C1-C6 alkoxy) carbonyl group, (C1-C6) alkoxy) carbonyl (C1-C6 alkyl) group, C1-C6 alkoxy group, trifluoro Methyl group, amino group, di (C1-C6 alkyl) amino (C1-C6 alkyl) group, acylamino group (formula:-NHCOC _n H2 _{n +1} , n is 0 to 6), -NH-CSC _n H _{2n + l} group (n is 0 to 6), terphenyl group, cyano group, C2-C6 alkynyl group, C2-C6 alkynyl group (substituted by C1-C6 alkyl or aryl group), hydroxy (C1-C6 alkyl) group, (C1- Independently from each other from a C6 acyl) oxy (C1-C6 alkyl) group, a (C1-C6 alkyl) thio group and an arylthio; Or R ₁ and R ₂ together may be a mono- or polycyclic C2-C20 alkylene group (optionally including at least one hetero atom except for 2,2 vsmethyltrimethylene group) or a C3--C12 cycloalkylene group Form.

R is selected from C1-C6 alkyl groups.

In the above definition, the aryl portion of an aryl group or arylalkyl group means an aromatic heterocyclic group such as an aromatic carbon-based group such as phenyl or naphthyl group or thienyl of a furyl group, and these functional groups are at least one selected from the group May have a substituent: a halogen atom, a C1-C4 alkyl group, a C1-C4 alkoxy group, a trifluoromethyl group, a nitro group and a hydroxyl group,

Oximes of 1,2-dithiol-3-thione derivatives such as those represented by the formula:

Additionally an aldehyde or ketone of the aforementioned compound, such as represented by the formula:

At least one compound represented by the following formula (A is C = N = OR ' ₃ and R' ₃ is an optionally substituted C1-C6 alkyl group (especially from hydroxyl, amino, chloro, bromo, fluoro, iodo Substituted with one or more functional groups selected) and a C 1 -C 4 alkoxy group or an aryl (C 1 -C 6 alkyl) group:

R ₃ has the same meaning as in the above definition.

One or more compounds represented by the formula (A is C = NO-CO-R '' ₃ , R '' ₃ is a t-hydrogen atom, an optionally substituted C1-C6 alkyl group, an aryl group and an aryl (C1-C6 alkyl) Term):

R '' ₃ Is a hydrogen atom, an optionally substituted C1-C6 alkyl group and an aryl group.

Another class of compounds in which A is a CH-OH group is formed, i.e. a compound represented by the formula:

Another class of compounds is formed wherein A is a functional group comprising C═N—R and is a Rdl C 1 -C 6 alkyl or aryl group, represented by the following formula:

Another class of compounds includes compounds represented by the following formula wherein A is a C═O group and X is an oxygen atom:

Where

R ₁ is hydrogen, halogen, nitro group, nitro group, thiocyano group, C1-C6 alkyl group, C2-C6 alkenyl group, aryl group, aryl (C1-C6 alkyl) group, aryl (C2-C6 alkenyl) group, Carboxyl group, (C1-C6 alkyl) carbonyl group, arylcarbonyl group, (C1-C6 alkoxy) carbonyl group, (C1-C6) alkoxy) carbonyl (C1-C6 alkyl) group, C1-C6 alkoxy group, trifluoromethyl group, amino group , Di (C1-C6 alkyl) amino (C1-C6 alkyl) group, acylamino group (formula:-NHCOC _n H2 _{n +1} , n is 0 to 6), -NH-CSC _n H _{2n + l} group (n is 0-6), terphenyl group, cyano group, C1-C6 alkynyl group, C2-C6 alkynyl group, C2-C6 alkynyl group (substituted by C1-C6 alkyl or aryl group), hydroxy (C1-C6 alkyl) group, (C1-C6 acyl) oxy (C1-C6 alkyl) group, (C1-C6 alkyl) thio group and arylthio group;

R ₂ is a nitro group, nitro group, thiocano group, C1-C6 alkyl group, C2-C6 alkenyl group, aryl group, aryl (C1-C6 alkyl) group, aryl (C2-C6 alkenyl) group, carboxyl group, (C1 -C6 alkyl) carbonyl group, arylcarbonyl group, (C1-C6 alkoxy) carbonyl group, (C1-C6) alkoxy) carbonyl (C1-C6 alkyl) group, C1-C6 alkoxy group, trifluoromethyl group, amino group, di (C1 -C6 alkyl) amino (C1-C6 alkyl) group, acylamino group (formula:-NHCOC _n H2 _{n +1} , n is 0 to 6), -NH-CSC _n H _{2n + l} group (n is 0 to 6) , Terphenyl group, cyano group, C2-C6 alkynyl group, C2-C6 alkynyl group (substituted by C1-C6 alkyl or aryl group), hydroxy (C1-C6 alkyl) group, (C1-C6 acyl) oxy (C1- C6 alkyl) group, (C1-C6 alkyl) thio group and arylthio group; or

R ₁ and R ₂ together form a mono- or polycyclic C ₂ -C 20 alkylene group and may optionally comprise one or more hetero atoms.

Another class of compounds includes one or more of the following compounds and pharmaceutically acceptable salts thereof:

R ₁ and R ₂ are hydrogen, halogen, nitro group, nitro group, thiocano group, C1-C6 alkyl group, C2-C6 alkenyl group, aryl group, aryl (C1-C6 alkyl) group, aryl (C2-C6 alkenyl ), Carboxyl, (C1-C6 alkyl) carbonyl group, arylcarbonyl group, (C1-C6 alkoxy) carbonyl group, (C1-C6) alkoxy) carbonyl (C1-C6 alkyl) group, C1-C6 alkoxy group, trifluoro Methyl group, di (C1-C6 alkyl) amino (C1-C6 alkyl) group, acylamino group (formula:-NHCOC _n H2 _{n +1} , n is 0 to 6), -NH-CSC _n H _{2n + l} group (n Is 0 to 6), terphenyl group, cyano group, C2-C6 alkynyl group, C2-C6 alkynyl group (substituted by C1-C6 alkyl or aryl group), hydroxy (C1-C6 alkyl) group, (C1-C6 acyl Or independently from each other selected from the group: oxy (C1-C6 alkyl), (C1-C6 alkyl) thio group and arylthio group; Or R ₁ and R ₂ together may be a mono- or polycyclic C2-C20 alkylene group (optionally including at least one hetero atom except for 2,2 vsmethyltrimethylene group) or a C3--C12 cycloalkylene group Form.

In the above definition, the aryl portion of an aryl group or arylalkyl group means an aromatic heterocyclic group such as an aromatic carbon-based group such as phenyl or naphthyl group or thienyl of a furyl group, and these functional groups are at least one selected from the group It may have a substituent: halogen atom, C1-C4 alkyl group, C1-C4 alkoxy group, trifluoromethyl group, nitro group and hydroxyl group. Isobenzothiazolone derivatives represented by one or more of the following structural formulas:

In the above structural formula, at least one of R ¹ and R ² is preferably nitro, arylazo, substituted arylazo, benzylideneamino or substituted benzylideneamino. If only one of R ¹ and R ² is substituted, R ¹ and R ² One of them is hydrogen. R ³ substituents are selected from alkyl groups having less than about 7 carbon atoms, amino, hydroxyl, alkoxyl and aryl groups (and functionalized forms thereof).

Preferred species of isobenzothiazole derivatives of the invention include, for example, derivatives in which R ¹ is arylazo and R ² is hydrogen. Exemplary compounds include R ² being hydrogen and R ¹ being phenylazo; Substituted phenylazo such as 4-hydroxyphenylazo; 4 nitro-2-methylphenylazo; 4 nitro-2-methylphenylazo; 2-hydroxy-1-naphthyl azo; 2-hydroxy-5-methylphenylazo; 2-hydroxy-4-methyl-5-nitrophenylazo; 4-hydroxy-1-naphthyl azo; 4-hydroxy-3-methyl-1-naphthylazo; 4-hydroxy-5-aza-1-naphthylazo; 2 amino-1-naphthyl azo; 1-hydroxy-2-naphthyl azo; 3-N, N dimethylaminopropylcarboxamido-1-hydroxy-4-naphthylazo: 1-hydroxy-4-methoxy-2-naphthylazo, 2-hydroxy-3-carboxy-1- Naphthyl azo; 1-hydroxy-3,6-disulfonate-2-naphthylazo: 2,3-dihydroxy-1-naphthylazo; Or 2-hydroxy-3,5-dimethyl-1-phenylazoin compound. In one embodiment, R ¹ is substituted benzylideneamino, 2,4-dinitrobenzylideneamino, and R ² is hydrogen. R ¹ is hydrogen, and R ² is 2-hydroxy-1-naphthyl azo or 4-hydroxy-1-phenylazo.

In one aspect, R ³ substituents with sufficient polarity can impart water solubility to the compound. For example, R ³ is — (CH ₂ ) nR ⁴ R ⁵ , where n is 2 to 6 and R ⁴ And R ⁵ is simple alkyls or hydrogen. Other possible water soluble side chains include 3-carboxypropyl, sulfonatoethyl and -ethyl ₂ of the type CH ₂ (CH ₂ OCH ₂ ) CH _3, where n is less than 10. Examples of preferred compounds are R ³ As side chains are aminoalkyl, carboxyalkyl, omega amino polyethylether and N-haloacetyl derivatives. In a broader sense, for a variety of uses, R ³ may be an alkyl, aryl, heteroaryl, alkoxy, hydroxyl or amino group, provided that R ³ is aminoalkyl, carboxyalkyl, hydroxy when including substituents for soluble or reactive purposes. It may be oxyalkyl or haloalkyl. R ³ The portion of aryl or heteroaryl can be substituted, for example, as aminoaryl, carboxyaryl or hydroxyaryl.

Also included are one or more isobenzothiazolones having the following structural formula:

Wherein at least one of R ¹ and R ² is nitro, arylazo, substituted arylazo, benzylideneamino or substituted benzylideneamino, and R ¹ and R ² Only one of hydrogen is R ³ is aminoalyl, aminoaryl and aminoheteroaryl, carboxyalkyl, carboxyaryl, or carboxyheteroaryl covalently linked to a polymer comprising an amino group or a hydroxy group. The spacer arm R ³ may comprise oligomers or polyethylene glycols and derivatives thereof. In one aspect, R ³ is 17-chloracetamido-3,6,9,12,15-pentaoxyheptadecyl and hexaethylene glycol is chloracetamide. When the polymer groups Y ¹ and R ³ contain a carboxyl group, it is preferable that the covalent bond is made through an ester bond. When the polymer contains an amino group, it is preferable that the analog covalent bond is made through an amide bond. The polymer with the amine may be a polymer such as chitosan, polyalkylamines, aminodextran, polyethyleneamine, polylysine or amitriene when coupled to R ³ .

The R ³ substituents of the invention may include alkyl linked to the polymer with the amine by substituting halogen with an amine in the alpha-haloalkyl or alpha-haloalkylcarboxamido R ³ precursor. In the case of aminoalkyl or aminoaryl groups, the R ³ substituent may be covalently linked to a polymer such as polyepichlorohydrin, chloromethylpolystyrene, polyvinylalcohol or polyvinylpyridine. The R ³ substituent of the present invention may generally be an aminoalkyl, hydroxyalkyl, aminoyl or hydroxyaryl group linked to a polymer comprising a carboxyl group via an amide or ester bond.

When the polymer is involved in the R ³ structure, the polymer is polyacrylic acid, polymethacrylic acid, polyitaconic acid, oxidized polyethylene oxide, poly (methylmethacrylate / methacrylic acid), carboxymethyl cellulose, carboxymethyl Such as agarose or carboxymethyl dextran. When such a carboxyl polymer is involved, R ³ can be aminoalkyl (eg 8 aminohexyl), hydroxyalkyl, aminoaryl or hydroxyaryl linked to the polymer via an amide or ester bond. In such cases, the R ³ precursor function is covalently attached to the polymer by reaction with the polymer with acid anhydride, or by curling with the polymer with carboxylate via carbodiimide-induced bond formation. Amine groups or hydroxyl groups to be linked may be provided.

In the compounds of the present invention, the R ³ substituent or precursor to it may be a haloalkyl or carboxyhaloalkyl moiety such as chloracetamido. Such substituents can be easily bonded to the polymer with the amine by substituting halogen with an amine.

In this specification, aryl is used with the intention of including organic residues derived from aromatic hydrocarbons or aromatic heterocyclic ring systems. Thus, aryl groups include unsubstituted ring residues such as phenyl and naphthyl and substituted forms thereof. The heterocyclic or heteroaryl moiety can be one that includes one or more heteroatoms (eg, N, O, S) in a ring system such as pyridyl, oxazolyl, quinolyl, thiazolyl, and substituted forms thereof. .

In this specification, alkyl is used with the intention to include aliphatic and cyclic organic moieties having carbon at the attachment site. Thus, alkyl groups include unsubstituted hydrocarbon moieties represented by the formula C _n H _{2n +1} and substituted and cyclic forms thereof. Such hydrocarbons are generally lower alkyls having less than six carbons. It is understood that larger alkyl groups may be used. Alkyl includes substituted moieties that include hydrocarbon moieties having one or more of the same or different functional groups described below.

The aforementioned alkyl group and aryl group may be substituted with a functional group. Such functional groups include all chemical groups which can be introduced by synthetic methods to produce stable compounds. Examples of such functional groups are hydroxyl, halogen (fluoro, chloro, bromo), amino (including alkylamino and dialkylamino), cyano, nitro, carboxy (including carboalkoxy), carbamoyl (Including N alkyl and N, N alkyl), sulfo, alcoholic, alkyl, aryl, arylazo and one or more of the following compounds:

R ₁ and R ₂ are independently (═O) or —OR, R is H or (C 1 -C 4) alkyl, and R ₃ is H or (C 1 -C 4) alkyl. Preferably R ₃ is H. Preferably, R ₁ and R ₂ are (= O) or -OR.

One or more of the following compounds are also included.

X represents H or two X's are a direct bond with two sulfur atoms; R ₁ is (═O) or —OH and R ₂ is H, Na, K or (C 1 -C 4) alkyl. In particular, the compound may be 3-keto lipoic acid, 3-hydroxy lipoic acid, 3-keto dihydrolipoic acid or 3-hydroxy dihydrolipoic acid.

1,2-dithiol-3-thione derivatives represented by the following formula:

R represents hydrogen, halogen, lower alkoxy group, lower alkyl group, amino group, lower alkyl substituted amino group or lower alkoxycarbonyl group. The term lower is used herein to mean isopropyl, isobutyl and tert-butyl as methyl, ethyl, propyl, and butyl and their structural isomers.

Preferred compounds among the compounds represented by the above formulas include the following compounds:

5- (4-phenyl-1,3-butadienyl) -1,2-dithiol-3-thione,

5-4 (4-chlorophenyl) -1,3-butadienyl-1,2-dithiol-3-thione,

5- {4 (4-methoxyphenyl) -1,3-butadienyl} -1,2-dithiol-3-thione,

5- {4- (p-toluyl) -1,3-butadienyl} -1,2-dithiol-3-thione,

5- {4- (o-chlorophenyl) -1,3-butadienyl} -1,2-dithiol-3-thione, and

5- {4- (m- (methylphenyl) -1,3-butadienyl} -1,2-dithiol-3-thione.

The following compounds are also included:

And 1,2-dithiol represented by the following formula:

Het is pyrimidin-2-yl, pyrimidin-2-yl, pyrimidin-4-yl, or pyrimidin-5-yl, or halogen, C1-C4 alkyl, C1-C4 alkoxy, mercapto, C1-C4 Alkylthio, said pyrimidin-2-yl, pyrimidin-2-yl, pyrimidin-4-yl, or pyrimidin-5-yl substituted by dialkylamino wherein each alkyl is C1-C4, and R is halogen, C1-C4 alkyl, C1-C4 alkyl substituted by alkoxycarbonyl with alkoxy C1-C4, carboxy, alkoxycarbonyl with alkoxy C1-C4, carbamoyl, N with alkyl C1-C4 -Alkylcarbamoyl, or R-CH (OH), where R represents hydrogen or alkyl of C1-C3.

N ⁶ Examples of benzyl adenosine or analogs, derivatives, metabolites, prodrugs or pharmaceutically acceptable salts thereof are described below.

In yet another embodiment, N ⁶ Benzyl Adenosine N ⁶ Benzyl adenosine-5'-monophosphate, which is described as a compound represented by formula (3). This compound has a molecular weight of 437.215 and a molecular formula of C ₁₇ H ₂₀ N ₅ O ₇ P.

(3)

In yet another embodiment, N ⁶ Benzyl Adenosine (N ⁶ -Benzyl) adenyl-p- (N ⁶ -benzyl) adenyl-p- (N ⁶ -benzyl) adenosine, which is a compound represented by the following formula (4). This compound has a molecular weight of 1373.39.

[Formula 4]

Carboxymethylcellulose (CMC) is a cellulose derivative having a carboxymethyl group bonded to a portion of the hydroxyl groups of glucopyranose that make up the cellulose backbone.

The active compounds disclosed herein can be prepared in the form of their pharmaceutically acceptable salts as can be seen in the above section. Pharmaceutically acceptable salts are salts that retain the desired biological activity of the parent compound while not imparting undesirable toxic effects. Examples of pharmaceutically acceptable salts are described in the literature (Berge et al., 1977, "Pharmaceutically Acceptable Salts," J. Pharm. Scl, Vol. 66, pp. 1-19).

The active compounds described herein may be prepared in the form of solvates. The term "solvate" is used herein to mean a complex of a solute (such as the active compound or salt of the active compound) and a solvent in the conventional sense. When the solvent is water, solvates may be referred to as hydrates, for example, hemihydrates, monohydrates, dihydrates, trihydrates, tetrahydrates, and the like.

The invention also includes prodrugs of the compounds of the invention, which can be converted into biologically active compounds by metabolism or hydrolysis. Prodrugs of any compound can be made using pharmacological techniques known to those of ordinary skill in the art.

Metabolites can be made after administration of a compound of the invention to a subject, eg through metabolism by molecular rearrangement or hydrolysis of the compound.

The present invention also encompasses the use of homologues and analogs of such compounds in addition to the urethra of the aforementioned compounds. As used herein, homologues are molecules having substantial structural similarity to the aforementioned compounds, and analogues are molecules having substantial biological similarity regardless of structural similarity.

 The invention also provides kits for carrying out the treatment regimens of the invention. Such kits may comprise, in one or more containers, a therapeutically effective amount of a composition of the present invention in a pharmaceutically acceptable form. Such kits may also further comprise instructions on how to use the compositions of the invention or the efficacy of the methods of the invention, and may further comprise information sheets for providing the physician with information appropriate for applying mucositis.

As used herein, the term "subject" means an animal, preferably a mammal, in particular a human. In an embodiment, a subject means a mammal, especially a human, as a human being or who will be exposed to radiation for radiotherapy, for example chemotherapy or radiotherapy.

The composition of the present invention is suitably administered parenterally. Parenteral administration can be intravenous or subcutaneous. In another embodiment, the route of administration may be rectal using suppository, transdermal or transmucosal administration.

In embodiments, compositions for the treatment and / or prevention of mucositis, weight loss and / or cachexia may be administered by topical application, including but not limited to oral and sublingual administration. Formulations suitable for topical administration include creams, gels, jellies, mucliages, pastes and ointments. In an embodiment, the composition may be formulated for transdermal administration, for example in the form of a transdermal patch.

An effective amount of a composition for the treatment and / or prevention of mucositis, and / or weight loss may be provided in a single dosage regimen or in a multiple dosage regimen.

In an embodiment, the composition may be administered orally, for example in the form of an oral rinse, or may be administered to the lungs as an aerosol via oral or nasal inhalation. In the case of administration via the oral or nasal inhalation route, the active ingredient is in a suitable pharmaceutical formulation and can be delivered using a mechanism in the form of a machine such as, for example, an inhaler or a medical nebulizer.

For intravenous injection, the active ingredient will be in the form of a parenterally acceptable aqueous solution, having no pyrogen and having the appropriate pH, isotonicity and stability. Methods for preparing appropriate solutions using isotonic vehicles, such as, for example, sodium chloride injections, Ringer's injections, and lactated Ringer's solutions, are known to those of ordinary skill in the art. If desired, preservatives, stabilizers, buffers, antioxidants and / or other additives may be included.

Pharmaceutical compositions for oral administration may be in tablet, capsule, powder or liquid form. Tablets may include solid carriers such as gelatin or adjuvant. Liquid phase pharmaceutical compositions generally comprise a liquid carrier such as water, petroleum, animal oil, vegetable oil, mineral oil or synthetic oil. Physiological saline, dextrose, or other sugar solutions or glycols such as ethylene glycol, propylene glycol or polyethylene glycol may be included.

Several delivery systems are known and can be used to administer the compositions of the present invention. In particular, the composition can be administered via microspheres, liposomes or other microparticulate delivery systems or sustained release formulations that are placed in specific tissues such as blood. Suitable examples of sustained-release carriers include semipermeable polymer matrices in the form of shared articles, such as, for example, suppositories or microcapsules. Injectable or microcapsule sustained release matrices such as polyactide are also provided.

Examples of the techniques and protocols mentioned above and other techniques and protocols that may be used in accordance with the present invention can be found in the literature (Remington's Pharmaceutical Sciences, 18th edition, Gennaro, AR, Lippincott Williams &Wilkins; 20th edition). (December 15, 2000) ISBN 0-912734-04-3 and Pharmaceutical Dosage Forms and Drug Delivery Systems; Ansel, HC et al. 7th Edition ISBN 0-683305-72-7. The entire contents set forth in this document are incorporated herein by reference.

Preferably, the compositions of the present invention are administered to a subject in a "therapeutically effective amount" as defined below. The actual amount and rate of administration and time course administered to an individual to achieve this effect will depend on the nature and severity of the condition being treated and factors such as the age, sex and weight of the patient being treated and the route of administration. . Toxicity and efficacy of such compositions can be determined by standard pharmaceutical methods.

Unless defined otherwise, all technical and scientific terms used herein have the meanings that are commonly understood by those of ordinary skill in the art.

Compounds described herein also include “other forms” of such compounds, including other forms that are protected by ionic, salt, solvate and substituents. For example, carboxylic acids (-COOH) include anionic (carboxylate) forms (-COO), salts or solvates thereof and conventional protected forms. Likewise, amino groups include conventional protected forms of amino groups. Hydroxyl groups likewise include anionic forms (-O-), salts or solvates thereof and common protected forms.

Certain compounds may exist in one or more specific geometric, optical, enantiomer, diastereomeric, epimeric, atropic, stereoisomer, tautomeric, conformational or anomer forms, for example cis -And trans-forms, E- and Z-forms, c-, t- and r-forms, endo and exo-forms, R-, S- and meso forms, 0- and L-forms, d- and 1- Forms, (+) and (-) forms, keno-, enol- and enolate-forms, syn and anti-forms, synclinal and anticlinal forms, alpha and beta forms, excalal and axial ( axial) and horizontal forms, boat-, chair-, twist-, envelop-, and half-chair-forms, and combinations thereof, collectively referred to herein as isomer or isomer forms.

Unless stated otherwise, the meaning of a particular compound includes all of the above isomeric forms, including (complete or partial) racemic mixtures or other mixtures thereof. Methods of preparing such isomeric forms (eg asymmetric synthesis) and separation methods (eg fractional crystallization and chromatography) are known in the art or may be readily used. Unless stated otherwise, the meaning of a particular compound is intended to include the ionic form, salt form, solvate form, and protected form of the compound.

The term "substituted" or "optionally substituted" means herein an unsubstituted or substitutable parent group. Unless stated otherwise, the term "substituted" is used herein in reference to a parent group having one or more substituents. The term "substituent" is used in the conventional sense and means a chemical moiety attached to the parent group or, where appropriate, bound to the parent group. A wide range of substituents are known, and their formation and introduction into various parent groups are also known to those of ordinary skill in the art.

Throughout the specification, the terms 'comprise' or 'include' or 'comprises', 'comprising', 'includes', unless the context otherwise requires. Or variations, such as 'including', is understood to include the specified integer, group of integers, and does not exclude other integers or groups of integers.

In this specification, "a, an" and "the" include both singular and plural forms unless the context clearly dictates otherwise. Thus, for example, "active substance" or "pharmaceutically active substance" includes not only a single active substance but also a combination of two or more different active substances, and the carrier is a single carrier as well as a combination of two or more carriers. Contains blends.

As used herein, the term "therapeutically effective amount" means an amount of the composition sufficient to have a beneficial effect on the subject. In particular, the beneficial effect may be the treatment, partial treatment or amelioration of one or more symptoms associated with mucositis. In the case of the prevention of mucositis, "therapeutically effective amount" means the amount of the composition necessary to prevent or inhibit the initial occurrence, progression or recurrence of mucositis or at least one symptom thereof.

As used herein, the terms “treatment” and related terms such as “treat” and “treating” reduce the progression, severity and / or duration of mucositis and ameliorate at least one or more symptoms of mucositis or reduce weight. To alleviate or prevent cachexia. Thus, the term "treatment" refers to any therapy that can have a beneficial effect on a subject. The treatment may be for a condition currently present or may be prophylactic (prophylactic treatment). Treatment may include therapeutic, ameliorating or prophylactic effects. As used herein, "therapeutic" and "prophylactic" treatments should be considered in the broadest sense. The term "therapeutic" does not necessarily mean treating the subject until the subject has fully recovered or until no weight loss or cachexia has occurred. Likewise, "prophylactic" does not necessarily mean that the subject will ultimately not develop mucositis or cachexia or will suffer from weight loss. Thus, therapeutic and prophylactic treatments include ameliorating the symptoms of mucositis and preventing or alleviating the risk of mucositis, cachexia and / or weight loss. The term "prophylactic" refers to alleviating the severity or incidence of mucositis, cachexia and / or weight loss, and the term "therapeutic" to be considered to alleviate the severity of currently existing mucositis, cachexia and / or weight loss. Can be.

The invention will now be described with reference to the following examples, which are provided for purposes of illustration only and are not intended to be construed as limiting the invention.

The present invention can be used to prevent and / or treat symptoms such as mucositis, weight loss, cachexia, etc., which are associated with radiation therapy or chemotherapy.

1 shows survival rates of rats when oral administration of 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione at different doses prior to exposure to 10 Gy gamma radiation Show Kaplan Meier's estimate.
FIG. 2 shows 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione for survival of mice exposed to 10 Gy gamma radiation at 1.33 Gy intensity per minute from a ⁶⁰ CO gamma radiation source. This is a bar chart detailing the effects of different doses.
FIG. 3 shows Kaplan Meier values for survival of rats when orally administered N ⁶ -isopentenyl adenosine at various doses prior to exposure to 10 Gy gamma radiation.
FIG. 4 shows N ⁶ -isopentenyl adenosine (6-gamma-dimethyl allyl aminopurine ribose (DAPR)) for survival of rats exposed to 10 Gy gamma radiation at 1.33 Gy intensity per minute from a ⁶⁰ CO gamma radiation source. This is a histogram detailing the effects of different doses.
FIG. 5 shows the 30-day survival rate of rats when various doses of 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione were previously administered and exposed to 8 Gy gamma radiation It is a graph.
FIG. 6 shows with or without treatment with 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione, and / or with or without exposure to gamma radiation. For the graph, a graph showing the change in body weight (as a percentage of the average body weight on the first day of treatment) of Swiss Albino rats.
FIG. 7 shows the 30-day survival rate of mice treated or not treated with 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione after exposure to gamma radiation at various doses. A graph representing.
Figure 8 shows radiation-induced micronuclei in rat bone marrow cells, with micro nucleated polychromatic erythrocytes indicated by arrows.
Figure 9 shows that radiation caused chromosomal aberrations in rat bone marrow cells, (a) normal metaphase showing 40 chromosomes, (b) chromatographic cleavage, exchange, And ring, (c) indicates powdering, and (d) indicates drainage.
FIG. 10 shows the average daily weight change of each animal and the mean for each treatment group of group (A) receiving a single treatment regimen and group (B) receiving a combination treatment regimen with radiation.
FIG. 11 shows the mean weight change as area under curve (AUC). AUC is calculated for the percentage of weight change seen in each animal in this study. This calculation was done using trapezoidal rule transformation. Group means were calculated and error bars represent SEM of each group. Groups were compared using one-way ANOVA. No statistically significant difference was observed between the group treated with 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione and the vehicle treated control group.
12 shows the average volume of tumors calculated by length and width measurements. Error bars represent SEM. (A) shows results for groups receiving only a single treatment regimen. (B) shows the results for the group receiving the combination therapy with radiation.
FIG. 13 shows mean weight change as area under the curve (AUC). AUC was calculated for the volume of tumor measured for each animal in this study. This calculation was calculated using a trapezoidal transformation. Group mean was calculated and error bars represent SEM of each group.

Example 1 Evaluation of the Radiation Protective Efficacy of 5- [2- pyrazinyl ] -4- methyl -1,2- dithiol- 3- thione (Oltifraz)

In this example, the efficacy and safety of 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione (oltipraz) as a radiation protection agent were evaluated.

Acute Toxicity Study

Animals are starved for 18 hours and then 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione (oltipraz) is added to 0, 100, 200, 400, 500, 600, 700, Observations were made for 14 days after administration in amounts of 800, 1000, 1250, 1500, 1750 and 2000 mg / kg.

Treatment group 1- CMC  And irradiation

Animals in Group 1 were orally administered 0.5% carboxymethylcellulose (CMC) prior to exposure to 10 Gy of gamma radiation.

Treatment group 2-5- [2- Pyrazinyl ]-4- methyl -1,2- Dithiol -3- Tion  And irradiation

Group 2 animals were treated with 5, 10, 25, 50, 100, 150 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione prior to exposure to 10 Gy of gamma radiation. Oral administration in an amount of 200 or 250 mg / kg.

Irradiation

One hour after administration of CMC or 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione, a cotton plug is placed into the restrainer in a specially designed ventilated acrylic box. The whole body was exposed to ⁶⁰ Co gamma radiation (Theratro, Atomic Energy Agency, Canada) with the animal lying down and fixed. Ten animals were irradiated at a dose of 1.33 Gy / min each time.

result

Acute Toxicity Study

In animals given different doses of 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione, there were no signs of any toxic effects up to 2 g / kg, 14 None of the animals died by the end of the day. Therefore, up to 2 g of 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione has been shown to be completely safe for administration. Further doses could not be evaluated due to drug dissolution issues.

Regarding the radioprotective efficacy of 5- [2-pyrazinyl] -4-methyl, rats were given 5- [2-pyrazinyl] -4-methyl-1,2-dithiol- prior to systemic exposure to 10 Gy gamma radiation. 3-thione was assessed by doses of 5, 10, 25, 50, 100, 150, 200 and 250 mg / kg body weight. After irradiation, animals were monitored daily for 30 days for symptoms of disease and death from radiation.

In the CMC and 10 Gy radiation exposure groups, symptoms of severe radiation diseases such as decreased food and water intake, irritability, drowsiness, weight loss, diarrhea, tearing, facial edema, weakness, and hair loss were induced. The first mortality was observed on day 4 in the CMC and irradiation groups, and all animals treated with radiation by 18 days after irradiation died.

Pretreatment of mice with various doses of 5- [2-pyrazinyl] -4-methyl delayed or alleviated the severity of symptoms of radiation disease. Incidence of radiation induced deaths was also delayed in the 5- [2-pyrazinyl] -4-methyl and irradiation groups compared to the CMC and irradiation groups. The longest delay was observed in the group treated with 100 mg / kg of 5- [2-pyrazinyl] -4-methyl, in which case the first death was observed on day 11 after irradiation (see Figure 1). This means that gastrointestinal symptoms are completely prevented. In contrast, the shortest delay was observed in the group treated with 5 mg / kg of 5- [2-pyrazinyl] -4-methyl, in which case the first death was observed on day 7 after irradiation (FIG. 1). Reference). This delay in mortality was also observed in other doses of the 5- [2-pyrazinyl] -4-methyl group.

Treatment of rats with varying doses of 5- [2-pyrazinyl] -4-methyl prevented damage to the gastrointestinal tract caused by radiation, which prevented 5- [2-pyrazinyl at all doses tested. ] Is evidenced by an increase in 10 day survival in the 4-methyl treated group (see FIG. 2). When 150 mg / kg and 200 mg / kg of 5- [2-pyrazinyl] -4-methyl was administered, no death occurred within 10 days after irradiation (see FIG. 2). The 30-day survival rate analysis showed that the survival of irradiated animals increased dose-dependently with 5- [2-pyrazinyl] -4-methyl up to 100 mg / kg, with CMC and irradiation not surviving mice. A survival rate of up to 60% was observed compared to the group (see FIG. 2). When compared to 100 mg / kg of 5- [2-pyrazinyl] -4-methyl and the irradiation group, increasing drug doses to 150 mg and 200 mg resulted in a 20% reduction in animal survival while 250 mg / kg. Increasing survival was found to be 30% (see Figure 2). Even at the lowest doses of 10, 25 and 50 mg / kg of the 5- [2-pyrazinyl] -4-methyl treatment group, survival rates were 20 and 30, respectively, compared to the CMC and irradiation groups that had no surviving rats. And 40% increase. Significant increases in survival were observed only in animals administered 50, 100, 150 and 200 mg of 5- [2-pyrazinyl] -4-methyl prior to exposure to 10 Gy (p> 0.05).

This example shows that orally administered 5- [2-pyrazinyl] -4-methyl protected mice from radiation induced disease and death. 100 mg compared to the other doses as it resulted in a 60% survival increase when compared to the irradiation group (control) that did not receive 5- [2-pyrazinyl] -4-methyl in the 100 mg / kg dose group. / kg was found to be the optimal dosage.

Example  2: N ⁶ - Isopentenyl  Evaluation of Radiation Protection Efficacy of Adenosine

In this example, the efficacy and safety of N ⁶ -isopentenyl adenosine (also known as 6-gamma-dimethyl allyl amino purine ribose (DAPR)) as a radiation protection agent were evaluated.

Acute Toxicity Study

Animals were starved for 18 hours and then observed for 14 days after DAPR was administered in amounts of 0, 100, 200, 400, 500, 600, 700, 800, 1000, 1250, 1500, 1750 and 2000 mg / kg.

Treatment group 1- CMC  And irradiation

Animals in Group 1 were orally administered 0.5% carboxymethylcellulose (CMC) prior to exposure to 10 Gy of gamma radiation.

Treatment group 2- DAPR  And irradiation

Group 2 animals were orally administered DAPR in amounts of 5, 10, 25, 50, 100, 150, 200 or 250 mg / kg body weight before exposure to 10 Gy of gamma radiation.

Irradiation

One hour after administration of CMC or DAPR, the whole body is ⁶⁰ Co gamma radiation (Theratro, Atomic Energy Agency) with a cotton plug inserted into a restrainer in a specially designed ventilated acrylic box with the animal lying down and fixed. , Canada). Ten animals were irradiated at a dose of 1.33 Gy / min each time.

result

Acute Toxicity Study

In animals given different doses of DAPR, no signs of toxic effects were seen, up to 2 g / kg, and no animals died until 14 days. Therefore, up to 2 g of DAPR appeared completely safe for administration. Further doses could not be evaluated due to drug dissolution issues.

Initially, 1, 5 and 10 mg / kg DAPR were also evaluated. However, there was no change in survival after irradiation. Therefore, subsequent evaluation experiments did not run this dose. The radiation protection efficacy of DAPR was assessed by administering DAPR in amounts of 0, 25, 50, 100, 150, 200 and 250 mg / kg body weight prior to systemic exposure to 10 Gy gamma radiation.

After irradiation, animals were monitored daily for 30 days for symptoms of disease and death from radiation. In the CMC and 10 Gy radiation exposure groups, symptoms of severe radiation diseases such as decreased food and water intake, irritability, drowsiness, weight loss, diarrhea, tearing, facial edema, weakness, and hair loss were induced. The first mortality was observed on day 4 in the CMC and irradiation groups, and all animals treated with radiation by 18 days after irradiation (see FIG. 3).

Pretreatment of mice with various doses of DAPR delayed or alleviated the severity of the symptoms of the radiation disease. Incidence of radiation induced deaths was also delayed in the DAPR and irradiation groups compared to the CMC and irradiation groups. The longest delay was observed in the group treated with 150 mg / kg of DAPR, in which case the first death was observed on day 11 after irradiation (see FIG. 3), indicating that gastrointestinal symptoms were completely prevented. . On the other hand, the shortest delay was observed in the group treated with 25 mg / kg DAPR, in which case the first death was observed on day 7 after irradiation (see FIG. 3). This delay in mortality was also observed in other doses of the DAPR group.

When mice were treated with varying doses of DAPR, damage to the gastrointestinal tract induced by radiation was prevented, evidenced by an increase in 10-day survival in the DAPR treated group at all doses tested (see FIG. 4). . When 150 mg / kg DAPR was administered, no death occurred within 10 days after irradiation (see FIG. 4). The 30-day survival rate increased the survival rate of irradiated animals up to 150 mg / kg, with DAPR dose-dependent increase, with up to 60% survival compared to the CMC and radiation groups without surviving rats. (See FIG. 4). Compared with the DAPR and irradiation groups of 150 mg / kg, increasing drug doses to 200 mg and 250 mg resulted in 20% and 30% reduction in animal survival (see FIG. 4). Even at the lowest dose of 25 mg / kg DAPR treatment group, survival was increased by 30% compared to the CMC and irradiation groups that did not have surviving mice. Significant increases in survival were observed only in animals that received 50, 100, 150 and 200 mg / kg of DAPR prior to exposure to 10 Gy (p> 0.05).

In this example, DAPR was completely safe up to this dose because no toxic side effects were observed up to 2 g / kg, and the mice were protected from radiation-induced disease and death in DAPR orally administered at all tested doses. Indicates. However, 150 mg / kg was found to be the optimal dose in terms of 60% increase in survival compared to other doses.

Example 3: of 5- [2- pyrazinyl ] -4- methyl -1,2- dithiol- 3- thione (oltipraz) Radiation protection effect

In this example, the efficacy and safety of 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione (oltipraz) as a radiation protection agent were evaluated.

animal

In this experiment, male musculus, 6-8 weeks old, weighing 25 ± 3 g from inbred colony (obtained from Hamadard University, Delhi) was used. Animals were kept under temperature and light controlled conditions in animal camps to randomly feed standard rat food (Hindustan Lever's Ltd., Delhi, India) and water.

Irradiation

For irradiation, a cobalt teletherapy unit (ATC-C9) was used at a cancer treatment center (Radiotherapy Department, SMS MEdical College & Hospital, Jaipur, India). The anesthetized animal was kept in a well-ventilated perspex box and the whole body was exposed to gamma radiation at a distance of 77.5 cm (SSD) from the radiation source to provide a dose of 1.33 Gy / min.

Acute Drug Toxicity

To measure the acute toxicity of oltipraz (5- [2-pyrazinyl] -4-methyl), animals were divided into four groups of ten animals and oltipraz (5- [2-pyrazinyl] -4-methyl -1,2-dithiol-3-thione) was orally administered for 2 days in an amount of 50, 100, 200, or 400 mg / kg body weight. Animals were continued for 30 days to determine mortality and other forms of toxic effects of oltipraz (5- [2-pyrazinyl] -4-methyl).

Determination of Optimal Dose of Oltipraz (5- [2-pyrazinyl] -4-methyl) for Radiation Protection

In order to select the optimal dose of oltipraz (5- [2-pyrazinyl] -4-methyl) for radiation protection, the animals were subjected to oltipraz (5- [2-pyrazinyl] -4-methyl- 1,2-dithiol-3-thione) was administered for 2 days in an amount of 50, 100, 200, or 400 mg / kg body weight / day. 30 minutes after the last dose, animals were exposed to 8 Gy gamma radiation. The survival of the animals was recorded for 30 days after irradiation. 30 minutes after exposure to radiation, reduced glutathione (GSH) and lipid peroxidation (LPO) levels in the liver and blood were assessed.

Reduced glutathione ( GSH ) analysis

The hepatic level of reduced glutathione (GSH) was measured according to standard methods. GSH in blood was measured with a spectrophotometer using Elman's reagent (DTNB) as a color developing agent. Absorbance at 412 nm was recorded using a UV-VIS Systronics spectrometer.

Lipid peroxidation ( LPO ) analysis

Lipid peroxidation levels in liver and serum were measured using Thiobarbituric Acid Reactive Substances (TABRS). Absorbance at 532 nm was recorded.

Dose reduction factor ( Dose Reduction Factor : DRF )

The protective performance of any agent (chemical or plant extract) is expressed as a dose reduction factor (DRF). Experimental animals by dividing the group of the LD _50/30 animals a group of LD _50/30 control may be DRF is calculated.

Control animal group (only radioactive)

Exposure to 6, 8, and 10 Gy gamma radiation was followed for 30 days to record mortality and radiation disease symptoms.

Experimental Animal Group ( Oltipraz (5- [2- pyrazinyl ] -4- methyl -1,2- dithiol- 3- thione )) and Irradiation

Animals in this group received 5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione for 2 days in an amount of 100 mg / kg body weight / day, the last dose It was then exposed to gamma radiation of 6, 8 and 10 Gy. In the same manner as the control animal group, animals were observed for 30 days for the mortality and radiation symptoms.

weight

The general condition and body weight were observed daily for all animal groups. For each group of rats, the percentage change in body weight was recorded daily by dividing by the average of the body weights of rats on the first day of treatment.

Endogenous spleen Colony  analysis

Endogenous spleen colony analysis was performed according to standard methods (Till and McCulloch). The endogenous spleen colony forming unit (CFU-S) was measured on day 10 after irradiation. Animals were killed by cervical dislocation. The spleens were separated, weighed and fixed with Bouin's fixative. Nodules visible to the naked eye on the surface of the spleen were visually counted. .

Survival rate analysis

Both groups (control and experimental groups) exposed to 6, 8, and 10 Gy gamma radiation were irradiated daily for 30 days, using the percentage of animals that survived 30 days after exposure for each radiation dose. Survival-dose response curves were created.

Quantitative changes in the spleen

To study the quantitative changes in the spleen, the spleens were weighed at each biopsy interval (day 1, 3, 7, 10, 14 and 30 days after irradiation).

Statistical analysis

The results obtained from the experiments are expressed as mean ± SE. The "t" test (Student's t test) was used for statistical comparison between groups. The significance level was P <0.01 and P <0.001. A regression analysis was performed to find the LD _50/30 levels and the dose reduction factor (DRF).

Analysis of Chromosomal Abnormalities

At the end of the experiment, chromosomal aberrations were analyzed for cytogenetic damage in bone marrow cells. Intraperitoneal injection (i.p.) of 0.026% colchicine for all animals was killed by cervical dislocation after 2 hours. Both femurs were dissected and separated. The metaface plate was prepared by air drying. The femoral bone marrow was aspirated, washed with physiological saline, treated with stock solution (0.6% sodium citrate), fixed with 3: 1 methanol: acetic acid, dried and stained with 4% Kimja (Giemsa: Sigma, USA). Chromosomal abnormalities were counted under light microscopy. A total of 400 metaface plates per animal were counted. Various chromosomal abnormalities were counted, such as chromatographic cleavage, chromosomal cleavage, fragmentation, cyclization, exchange, and two conjugation. When both chromatids have a cleavage, it is called a chromosome type abnormality, and the chromatized type abnormality was only the case where only one chromatized abnormality existed. If the nodule is not clearly associated with a particular chromosome, it is called a fragment.

result

Radiation Protection Efficacy of Oltipraz (5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione) on Radiation-induced Diseases, Weight Changes, Spleen Colonies and Animal Survival Was studied using Swiss Albino rats. No toxic effects occurred in mice treated with oltipraz (5- [2-pyrazinyl] -4-methyl-1,2-dithiol-3-thione) for two consecutive days. Rather, these rats showed an increase in body weight at 30 days compared to sham irradiated (normal) animals.

The optimal dose of oltipraz (5- [2-pyrazinyl] -4-methyl), which shows the maximum radiation protection effect, is 100 mg / kg body weight per day for two consecutive days before irradiation. Appeared (see FIG. 5).

No significant changes in GSH concentrations in liver and blood were observed in normal and olifraz (5- [2-pyrazinyl] -4-methyl) treated animals (Table 2). However, a significant decrease in GSH concentration was observed in the control animal group (only radiation treatment), whereas in the experimental animal group various oligraz (5- [2-pyrazinyl] -4 groups were compared with the control animal group. -Methyl) concentration showed a significant increase in GSH concentration (concentration in liver and blood) (Table 2). Maximum increase in GSH concentration was observed in the group of animals pretreated with olifraz (5- [2-pyrazinyl] -4-methyl) in an amount of 100 mg / kg body weight / day.

Increases in TBARS levels in liver and serum were evident in the control group compared to normal animals, and this was seen between normal and oltipraz (5- [2-pyrazinyl] -4-methyl) treated animals. No significant difference was observed in the levels. Significant dose dependent decreases were observed in the pretreated and irradiated groups of oltipraz (5- [2-pyrazinyl] -4-methyl). However, the maximum reduction in LPO levels was measured in the group of animals pretreated with olifraz (5- [2-pyrazinyl] -4-methyl) in an amount of 100 mg / kg body weight / day.

In this study, pretreatment with oltipraz (5- [2-pyrazinyl] -4-methyl) was observed to improve survival of mice exposed to various doses of gamma radiation (see FIG. 5). Signs of radiation diseases such as drowsiness, diarrhea, weight loss, matted hair, hair loss, facial edema and loss of appetite have been observed in animals exposed to various doses of gamma radiation (control group). The severity of the radiation disease is dose dependent, with 38% of animals dying within 30 days if irradiated at 6 Gy, and mortality rates at 14 and 10 days in the control animal group, respectively, when exposed to 8 Gy and 10 Gy. Was observed to be 100% (see FIG. 7). No radiation speculation was observed in the group of animals treated with 100 mg / kg body weight / day of olifraz (5- [2-pyrazinyl] -4-methyl) prior to exposure to 6 Gy. However, after exposure to 8 Gy and 10 Gy radiation the severity of radiation vaginality was much less compared to the respective controls. Survival in the 6 Gy experimental group was 100%, but the survival rate was reduced to 61% and 20% in the 8 Gy and 10 Gy experimental group, respectively (see Figure 7).

Regression analysis of the survival data showed that LD _50/30 was 6.24 Gy and 8.82 Gy for control animals and experimental driving groups, respectively. On the basis of LD _50/30 DRF value was calculated to be 1.25.

The maximum weight loss was 24% and the minimum weight loss was 13.5% in the control group, compared to 22.5% and 1.7% in each experimental group. In addition, the experimental animals showed body weight gains of 17% (6 Gy), 9.5% (8 Gy) and 13.7% (10Gy) on the 30th day from the time of irradiation. .

Endogenous spleen colony analysis and spleen weight change analysis were performed to evaluate the protective effect of oltipraz (5- [2-pyrazinyl] -4-methyl) on radiation damage to liver tissue. Mice pretreated with oltipraz (5- [2-pyrazinyl] -4-methyl) significantly increased the number of spleen colonies compared to mice treated with radiation alone (Table 3). The pattern of spleen weight change was similar for all control animals (radiation only) until day 7 after irradiation, but the decrease in spleen weight was dose dependent, i.e. the weight loss increased with higher radiation dose. . Maximum weight loss was observed on day 7 after the increase in tissue weight was recorded. In addition, an increase in the weight of the spleen was observed, which was greater than normal on day 14 and reached normal levels on day 30 in animals treated with 6 Gy. Animal groups were unable to survive for more than 14 days (8 Gy), and 10 days (10 Gy) in the group of animals exposed to radiation (see FIG. 7). In the animals treated with oltipraz (5- [2-pyrazinyl] -4-methyl) and radiation (group of experimental animals), the weight of the spleen decreased until the 7th day, but this decrease was achieved at each biopsy interval. Significantly smaller compared to After 7 days, a gradual increase was observed and reached almost normal levels by 30 days.

Chromosome research

Oral administration of 5- [2-pyrazinyl] -4-methyl (100 mg / kg body weight / day) prior to exposure to gamma radiation has shown that chromosomal damage is effectively prevented in Swiss albino mice (FIG. 9). , Table 5 and Table 6). Animals exposed to 8 Gy gamma radiation showed chromosomal damage in the form of chromatographic cleavage, chromosomal cleavage, isotopic rings, two isotopes, exchange and fragments without a centrio. After 6 hours of irradiation, the frequency of abnormal cells increased significantly. Maximal abnormal cells were observed 12 hours after biopsy. In addition, the frequency of aberrant cells was found to be reduced in late biopsy events after irradiation. However, the animals pretreated with 5- [2-pyrazinyl] -4-methyl significantly reduced the frequency of aberrant cells compared to the control group treated with radiation. The number of micronuclei was significantly increased in 8 Gy radiation treated mice. However, the number of micronuclei was significantly reduced in the group of animals pretreated with 5- [2-pyrazinyl] -4-methyl (see Table 4 and FIG. 8).

Table 1: Radiomodulation Effect of 5- [2-pyrazinyl] -4-methyl on 30-day Viability of Swiss Albino Rats

Table 2: Radiomodulation Effects of 5- [2-pyrazinyl] -4-methyl on GSH and LPO Levels in Liver and Blood of Swiss Albino Rats

Significance level: ^a p <0.05, ^b p <0.005, ^c p <0.001

Table 3: Spleen Responses on Day 10 after Irradiation with or without 5- [2-pyrazinyl] -4-methyl in Swiss Albino Rats

Level of significance: ^a p <0.05, ^b p <0.005, ^c p <0.001. ns = no survivors

Table 4: Frequency of Micronuclei of Bone Marrow Cells of Swiss Albino Rats with or without 5- [2-pyrazinyl] -4-methyl Treatment After 8 Gy Gamma Irradiation

Each figure averages ± SE

Control group = 8.0 Gy gamma-ray

Experimental group = 5- [2-pyrazinyl] -4-methyl + 8.0 Gy gamma ray

Normal = do not process

Level of significance: ^a p <0.05, ^b p <0.005, ^c p <0.001.

Table 5: Table 4: Frequency of chromosomal abnormalities in 5- [2-pyrazinyl] -4-methyl treated or untreated Swiss albino rats after 8 Gy gamma irradiation

Each figure has an average of ± SE. A total of 400 metafaces are measured per animal.

Level of significance: ^a p <0.05, ^b p <0.005, ^c p <0.001.

Table 6: Frequency of Chromosome Aberrations in 5- [2-pyrazinyl] -4-methyl Treated or Untreated Swiss Albino Rats After 8 Gy Gamma Irradiation

Each figure has an average of ± SE. A total of 400 metafaces are measured per animal.

Level of significance: ^a p <0.05, ^b p <0.005, ^c p <0.001.

Example 4: 5- [2- pyrazinyl ] -4- methyl -1,2- dithiol- 3-thione (oltipraz) for the prevention of weight loss and reduction of tumor growth in combination with radiation therapy evaluation

The purpose of this study was to evaluate the efficacy of 5- [2-pyrazinyl] -4-methyl (oltipraz) in inhibiting tumor growth and preventing weight loss. NCI H146 in nude mice A small cee lung cancer model was used and combined with monotherapy and radiation therapy.

Study design

96 female nude mice ( nu ⁺ / nu ⁺ ) were randomly divided into 8 treatment groups. The lower left flank of each rat was inoculated with 0.05 mL of 1 × 10 ⁶ NCI-H146 small cell lung cancer cells with matrigel. Treatment started when the tumor reached a volume of 75-125 mm ³ . For each group the vehicle, radiation, 5- [2-pyrazinyl] -4-methyl was treated as detailed in Table 7.

Table 7: Study Design

The start date of drug treatment was designated as day 1. Carriers were administered orally once a day on Days 1 and 3 for rats in Groups 1 and 5. For rats in groups 2, 3, 6, and 7, 5- [2-pyrazinyl] -4-methyl (50 mg / kg or 100 mg / kg) with the carrier once daily on day 1 and day 3 Administration to the gastrointestinal tract through the oral cavity. For rats in groups 4 and 8, 5- [2-pyrazinyl] -4-methyl (50 mg / kg) was administered to the gastrointestinal tract with the vehicle once daily, from day 1 to day 20. Rats in groups 5-8 were treated with radiation. Irradiation was performed twice at 2 Gy / dose on day 2 and day 4. These experiments were anesthetized with ketamine (100 mg / kg) and xylazine (5 mg / kg) in these groups of rats and then placed under a radiation shield so that the tumor portion of the flank was exposed to radiation. The radiation was irradiated using a Philips 160 kV source at a dose rate of about 1.0 Gy / min at a focal length of about 40 cm. Tumors were measured every other day during the study. On day 21, all mice were killed and the remaining tumors were excised, weighed, photographed and fixed in formalin solution for subsequent analysis.

Weight measurement and survival rate

In order to analyze the difference in body weight of the animals in the treatment group as a marker of possible toxicity that may result from the treatment, daily weighting and survival rates were recorded for all animals. Animals showing 20% or more weight loss compared to the body weight at the start of the study were euthanized during the study period.

Tissue culture

H146 human lung cancer cells were obtained from ATCC. These cells were cultured in DMEM supplemented with 10% fetal calf serum (FCS), penicillin, streptomycin and 2 mM L-glutamine. The medium was removed and washed twice with calcium and sterile magnesium-free phosphate buffered saline (PBS) and cells were passaged by adding 1-2 ml of 0.25% trypsin and 0.03% EDTA solution. The flask was left at 37 ° C. until the cells detached. The cells were then subcultured at a ratio of 1: 3.

Place of study

The study was conducted at a Biomodels 5- [2-pyrazinyl] -4-methylLAC certified facility in Watertown, Massachusetts. IACUC clearance for this study was obtained from Biomodels IACUC.

animal

As experimental animals, and the weeks 5-6 weeks, before treatment the average weight is 23.8 g, isoforms for genes nu (nu ⁺ / nu ⁺⁾ in female nude mice (Charles River Labs, strain code 088 ; Crl-NUFoxn1 ^nu ). Holes in the ears were numbered individually for each animal and housed in groups of 5-6 per cage. The animals were adapted before beginning the study. During this period of at least two days animals were observed daily to remove animals that were in poor condition. Nude mouse colonies from Charles River Labs were established using mice obtained from NIH derived from naturally occurring mutations resulting in complete lack of thymic epithelium and significant reduction of hair and whiskers. Lack of thymic epithelium prevents maturation of T-cells, resulting in a significant deficiency of cell mediated immune responses. Such animals are generally considered immunodeficiency animals and are sensitive to non-syngenic tumors.

housing

This study was conducted in an animal room provided with filtered air at a temperature of 70 ° F ± 5 ° F and a relative humidity of 50% ± 20%. This animal room was set up to maintain at least 12-15 air changes per hour. An automatic timer was installed in the animal room for a light / dark cycle with 12 hours of "on" and 12 hours of "off" without Tweetlight.

Sterile Bed-O-Cobs® bedding was used. This bedding was replaced at least once a week.

Animal cages, tops, bottles and the like were formulated with commercially available detergents and dried in air. These were wrapped and autoclaved prior to use. Commercially available sterilizers were used to sterilize the materials and surfaces introduced into the hood. Floors were wiped daily and wiped with commercially available detergent at least twice a week. Walls and cage shelves were cleaned at least once a month with diluted bleach solution. All cages were marked with a cage card or label containing the required information to identify the project name, dose, animal number, and treatment group. Temperature and relative humidity were recorded and recorded during the study.

Diet

Animals were fed sterile feed (Labdiet® 5053 (pre-sterilized) rodent chow) and sterile water optionally supplied.

Random Classification and Allocation of Animals

Prior to starting treatment, animals were optionally divided into eight treatment groups. Each animal was punched into the ear according to an individual number for identification. To identify each cage, a cage card was used to indicate study number (CAN-01), treatment group number, and animal number.

Evaluation of the results

Statistical differences between treatment groups were determined by the following analysis with a critical value of 0.05: Student's t-test, Mann-Whitney U test, and chi-square analysis.

Experimental Method

Using a micro-potassium Perth measuring tumor size every two days and the tumor volume was calculated (length x width) ³ as Π / 3. Tumor growth index (TGI) was calculated using the formula (100- (Vc ^* 100 / Vt): Vc is the average volume of tumors in the control group and Vt is the average volume of tumors in the experimental group). .

result

All four animals were observed dead in this study. Three occurred with anesthesia used to immobilize the animals for radiation treatment (two occur on day 2 in groups 6 and 7 and one on day 4 in group 7). The fourth death occurred at day 15 in the 100 mg / kg 5- [2-pyrazinyl] -4-methyl treatment group on radiation plus day 1 and day 3.

Weight loss (FIGS. 10 and 11)

The average daily weight change rate for each treatment group is shown in FIG. 10. Weeks in the control group showed an average weight gain of 1.8% compared to body weight at the start of the experiment by Day 21. Mice treated with 50 mg / kg of 5- [2-pyrazinyl] -4-methyl on Day 1 and Day 3 showed an average 0.1% weight loss compared to body weight at the start of the experiment by Day 21. Mice treated with 100 mg / kg of 5- [2-pyrazinyl] -4-methyl on Day 1 and Day 3 showed an average weight gain of 4.8% compared to body weight at the start of the experiment by Day 21. Mice treated with 50 mg / kg of 5- [2-pyrazinyl] -4-methyl from Day 1 to Day 20 showed an average weight gain of 0.8% compared to body weight at the start of the experiment until Day 21. Rats treated with radiation and vehicle showed an average weight gain of 2.6% compared to body weight at the start of the experiment by Day 21. Rats treated with radiation and 50 mg / kg of 5- [2-pyrazinyl] -4-methyl on Day 1 and Day 3 showed an average weight gain of 0.8% compared to body weight at the start of the experiment by Day 21. Rats treated with 100 mg / kg of 5- [2-pyrazinyl] -4-methyl on day 1 and day 3 showed an average weight gain of 5.1% compared to body weight at the start of the experiment by Day 21. Rats treated with radiation and 50 mg / kg of 5- [2-pyrazinyl] -4-methyl from Day 1 to Day 20 showed an average weight gain of 1.4% compared to body weight at the start of the experiment by Day 21.

The significance of these differences was assessed by calculating the area under the curve (AUC) for the mean weight change for each animal and then comparing the groups using the One-Way ANOVA test. There was no significant difference between the 5- [2-pyrazinyl] -4-methyl treated group and the vehicle control group (P = 0.153). Radiation treated + group treated with 100 mg / kg of 5- [2-pyrazinyl] -4-methyl on Day 1 and Day 3 and radiation treatment + 5- [2-pyrazinyl from Day 1 to Day 20 ] There was a significant difference between the groups treated with 100 mg / kg of 4-methyl (P = 0.003). AUC data is shown in FIG. 11.

Tumor Volume (Figure 12)

From the measurements of length and width measured every other day, the average radius (r), which is the sum of length and width divided by 4, was calculated, and the tumor volume was calculated using Equation 4/3 πr ³ . Data for average tumor volume is shown in FIG. 12. The mean tumor volume for the vehicle control group increased from 109 mm ³ on Day 1 to 1374 mm ³ on Day 21. The mean tumor volume for the groups treated with 100 mg / kg of 5- [2-pyrazinyl] -4-methyl on Day 1 and Day 3 increased from 110 mm ³ on Day 1 to 1341 mm ³ on Day 21 It was. The mean tumor volume for the groups treated with 5- [2-pyrazinyl] -4-methyl 50 mg / kg from Day 1 to Day 20 ranged from 76 mm ³ on Day 1 to 1130 mm ³ on Day 21 Increased. Mean tumor volume for the radiation plus carrier control group increased from 92 mm ³ on Day 1 to 339 mm ³ on Day 21. The mean tumor volume for the groups treated with 50 mg / kg of 5- [2-pyrazinyl] -4-methyl on radiation plus Day 1 and Day 3 ranged from 93 mm ³ on Day 1 to 971 mm on Day 21 Increased to ^three . The mean tumor volume for the group treated with 100 mg / kg of radiation plus 5- [2-pyrazinyl] -4-methyl on Day 1 and Day 3 ranged from 63 mm ³ on Day 1 to 769 mm on Day 21 Increased to ^three . The mean tumor volume for the groups treated with radiation treatment plus 5- [2-pyrazinyl] -4-methyl 50 mg / kg from Day 1 to Day 20 ranged from 140 mm ³ on Day 1 to 1380 on Day 21 increased to mm ³ .

Further analysis of the experimental data was performed by calculating the area under the curve (AUC) for tumor volume for each animal and comparing the data between groups using the One-Way ANOVA (ranks test). Overall analysis did not show a significant difference between the 5- [2-pyrazinyl] -4-methyl treated group and the carrier control group (P = 0.052). However, the Mann-Whitney Rank sum test showed that there was a significant difference between the radiotherapy + carrier group and the carrier-only group (P = 0.004). There was also a significant difference between the group treated with only the carrier and the group treated with 50 mg / kg of 5- [2-pyrazinyl] -4-methyl on Days 1 and 3 (P = 0.030). Obviously, at least in the dosages and dosing schedules used in this study, 5- [2-pyrazinyl] -4-methyl has no additional effect when treated in combination with radiation therapy, but 5- [2-pyra Genyl] -4-methyl appears to be effective as a single agent. To assess the effect of 5- [2-pyrazinyl] -4-methyl on radiation therapy, comparisons were made using ANOVA (rank analysis) on the radiation treated groups. No significant difference was observed between the radiation treated group and the radiation treated + 5- [2-pyrazinyl] -4-methyl treated group (P = 0.177). Analysis using the Mann-Whitney rank sum test showed no statistically significant difference between groups (50 mg / kg of 5- [2-pyrazinyl] -4-methyl on Days 1 and 3) P = 0.012 for groups; P = 0.977 for groups treated with 5- [2-pyrazinyl] -4-methyl 100 mg / kg on Days 1 and 3; 5 from Day 1 to Day 20 P = 0.012) for groups treated with 50 mg / kg of [2-pyrazinyl] -4-methyl. Tumor volume AUC data is shown in FIG. 13.

Discussion

In this study, we tested the efficacy of 5- [2-pyrazinyl] -4-methyl in inhibiting tumor growth and reducing weight loss during radiation therapy using a rat NCI-H146 small cell lung cancer model. Carrier (0.5% aqueous CMC solution), radiation alone (2 doses of 2 Gy on Day 2 and Day 4), rats with tumors on Day 1 and Day 3, or Day 1 for rats with tumors Various treatments were performed by a combination of 50 [mg] / kg or 100 mg / kg of 5- [2-pyrazinyl] -4-methyl, or radiation treatment plus 5- [2-pyrazinyl] -4-methyl treatment by day 20. . 5- [2-pyrazinyl] -4-methyl showed no evidence of toxicity in this study as judged from survival and weight change data. As a single agent, 5- [2-pyrazinyl] -4-methyl was effective in reducing tumor growth, and 50 mg / kg of 5- [2-pyrazinyl] once daily on Days 1 and 3 Administration of -4-methyl resulted in a significant decrease in tumor volume compared to the carrier control group on Day 21. As expected, treatment with radiation alone was effective in reducing H146 tumor growth (P = 0.004). There was no statistically significant difference between the group receiving radiation alone and the group receiving radiation plus 5- [2-pyrazinyl] -4-methyl (P = 0.177).

All documents cited herein are hereby incorporated by reference. Various modifications and variations to the embodiments of the invention described above will be apparent to those skilled in the art without departing from the scope of the invention. Although the invention has been described with reference to specific preferred embodiments, the invention as claimed should not be unduly limited to these specific embodiments. Indeed, various modifications to the methods of carrying out the invention described above, which will be apparent to those skilled in the art, are intended to be included in the invention. References to prior art documents herein are not to be taken in a form that admits or suggests that these prior art documents form part of the general general knowledge in any of the countries.

Claims (4)

N ⁶ - iso-pentenyl adenosine or N ⁶ - benzyl adenosine, or a pharmaceutically acceptable salt thereof or mucositis therapeutic to prevent pharmaceutical composition comprising a solvate and a pharmaceutically acceptable carrier. The method of claim 1,
Pharmaceutical composition, characterized in that the composition further comprises a chemotherapeutic agent. The method according to claim 1 or 2,
Pharmaceutical composition, characterized in that the composition further comprises carboxymethylcellulose. The method of claim 1,
The mucositis is a pharmaceutical composition, characterized in that caused by one or more of chemical damage, biological damage, and damage by radiation.

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